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CRISPR header icon CRISPR Plasmids - Double-Strand Break (Cut)


Fully functional Cas9 enzymes designed to introduce a double-strand break (DSBs) at a specific location based on a co-expressed gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ); a mechanism which frequently causes insertions or deletions (InDels) in the DNA, possibly resulting in frameshifts. If a repair template with high homology to the DNA surrounding the DSB is introduced along with the Cas9 and gRNA plasmids, the cell may instead repair the break using homology-directed repair (HDR). HDR is much less error-prone than NHEJ and can be used to faithfully introduce specific genomic changes.



Mammalian

Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
hCas9Cas9CMV Church RNA-Guided Human Genome Engineering via Cas9. Science. 2013 Jan 3. Expresses human codon optimized Cas9 nuclease for genome engineering pcDNA3.3-TOPO
pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-purohumanized S. pyogenes Cas9Puromycin Zhang Multiplex Genome Engineering Using CRISPR/Cas Systems. Science. 2013 Jan 3. This plasmid separately encodes a human codon-optimized SpCas9, a tracrRNA and customizable crRNA. pUC ori vector
pX330-U6-Chimeric_BB-CBh-hSpCas9humanized S. pyogenes Cas9CBh Zhang Multiplex Genome Engineering Using CRISPR/Cas Systems. Science. 2013 Jan 3. A human codon-optimized SpCas9 and chimeric guide RNA expression plasmid. pUC ori vector
  • Tag / Fusion Protein
    • 3xFLAG (N terminal on insert)
  • pMJ920Cas9 (Synthetic)CMV Doudna RNA-programmed genome editing in human cells. elife. 2013;2:e00471. doi: 10.7554/eLife.00471. Epub 2013 Jan 29. MacroLab 6D
    JDS246mammalian codon-optimized streptococcus pyogenes Cas9 - 3X FlagCMV Joung Joung lab unpublished CRISPR plasmids (unpublished) Expresses mammalian codon optimized Cas9 nuclease with C-term 3X FLAG from CMV and T7 promoters unknown
    p3s-Cas9HCCas9CMV Kim Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease. Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2507. pCDNA3.1
    pCas9_GFPCas9-2A-GFP (Synthetic)CAG Musunuru Cas9 GFP plamsids (unpublished) Co-expression of human codon-optimized Cas9 nuclease and GFP, plasmid optimized for expression in human pluripotent stem cells pCAG
    pST1374-NLS-flag-linker-Cas9Human codon optimized Cas9CMVBlasticidin Huang Generation of gene-modified mice via Cas9/RNA-mediated gene targeting. Cell Res. 2013 Apr 2. doi: 10.1038/cr.2013.46. pST1374 (Addgene #13426)
    pSimpleII-NLS-NmCas9-HA-NLS(s)NmCas9 (Other)EF1a Thomson Efficient genome engineering in human pluripotent stem cells using Cas9 from Neisseria meningitidis. Proc Natl Acad Sci U S A. 2013 Aug 12. This plasmid encode NmCas9 with two NLS and a HA tag pSimpleII
    pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s)NmCas9 (Other), U6pr-tracrRNA (Other)EF1a, U6 Thomson Efficient genome engineering in human pluripotent stem cells using Cas9 from Neisseria meningitidis. Proc Natl Acad Sci U S A. 2013 Aug 12. This plasmid contains expression cassette for NmCas9 with N and C NLS and an HA tag, a cassette for expression of tracrRNA, a cassette for cloning crRNA under the control of U6 promoter. pSimpleII
    pSimpleII-NmCas9-FLAGNmCas9 (Other) Thomson Efficient genome engineering in human pluripotent stem cells using Cas9 from Neisseria meningitidis. Proc Natl Acad Sci U S A. 2013 Aug 12. FLAG tagged NmCas9 with no NLS. pSimpleII
    pSpCas9 (PX165)hSpCas9 (Synthetic)Cbh Zhang Genome engineering using the CRISPR-Cas9 system. Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24. Human codon optimized Cas9 nuclease from Streptococcus pyogenes (Cbh-3X-FLAG-NLS-SpCas9-NLS) PX165
    pSpCas9(BB)-2A-GFP (PX458)hSpCas9 (Synthetic)Cbh Zhang Genome engineering using the CRISPR-Cas9 system. Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24. Cas9 from S. pyogenes with 2A-EGFP, and cloning backbone for sgRNA PX458
    pSpCas9(BB)-2A-Puro (PX459)hSpCas9-2A-Puro (Synthetic)CbhPuromycin Zhang Genome engineering using the CRISPR-Cas9 system. Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24. NOTE: A new version of this plasmid is now available. See Addgene plasmid 62988. PX459
    pCAG-T3-hCAS-pACodon optimized Cas9 (Synthetic)CAG promoter Fujii Efficient generation of large-scale genome-modified mice using gRNA and CAS9 endonuclease. Nucleic Acids Res. 2013 Aug 30. Expresses CAS9 nuclease in mammalian cells. Used to modify genome of mouse embroys pCAGGS
    M-SPcasCas9 (Other)CMVNeomycin (select with G418) Church Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. Mammalian S. pyogenes Cas9 expression, human optimized pcDNA3.3 TOPO
    M-ST1casCas9 (Other)CMVNeomycin (select with G418) Church Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. Mammalian S. thermophilus #1 Cas9 expression, human optimized pcDNA3.3 TOPO
    M-NMcasCas9 (Other)CMVNeomycin (select with G418) Church Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. Mammalian N. meningitidis Cas9 expression, human optimized pcDNA3.3 hygro
    pCW-Cas9humanized S. pyogenes Cas9Tet ONPuromycin Sabatini Genetic screens in human cells using the CRISPR-Cas9 system. Science. 2014 Jan 3;343(6166):80-4. doi: 10.1126/science.1246981. Epub 2013 Dec 12. Doxycycline-inducible lentiviral expression of SpCas9 pCW57.1
    pCAG-hCas9hCas9CAGNeomycin (select with G418) Hatada Generation of an ICF syndrome model by efficient genome editing of human induced pluripotent stem cells using the CRISPR system. Int J Mol Sci. 2013 Sep 30;14(10):19774-81. doi: 10.3390/ijms141019774. Expresses hCas9 under the CAG promoter for CRISPR pEGFP-N1
    lentiCRISPR v2Cas9 (Synthetic), Puromycin resistanceEFS-NS, EFS-NSPuromycin Zhang Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047. Replaces original lentiCRISPRv1 (Addgene Plasmid 49535) and produces ~10-fold higher titer virus. 3rd generation lentiviral backbone. Custom
    lentiCas9-BlastCas9 (Synthetic), Blasticidin resistanceEFS-NS, EFS-NSBlasticidin Zhang Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047. Expresses human codon-optimized S. pyogenes Cas9 protein and blasticidin resistance from EFS promoter. 3rd generation lentiviral backbone. pFUGW
    pLenti-OC-IRES-BSDOCT1 (Homo sapiens), Cas9 (Other)CMVBlasticidin Wei High-throughput screening of a CRISPR/Cas9 library for functional genomics in human cells. Nature. 2014 Apr 9. doi: 10.1038/nature13166. To create stable cell clones with high-level expression of Cas9 and OCT1 pLenti-CMV-BSD POU2F1 OCT1, OTF1, oct-1B
    pLV hUbC-Cas9-T2A-GFPhumanized Cas9 T2A GFP (Other)hUbC Gersbach Multiplex CRISPR/Cas9-based genome engineering from a single lentiviral vector. Nucleic Acids Res. 2014 Aug 13. pii: gku749. 3rd generation transfer vector. Co-expresses human optimized S. pyogenes Cas9 and GFP FUGW
    pSQT834Csy4-T2A-Cas9-NLS (Homo sapiens)CAG Joung Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. Nat Biotechnol. 2014 Apr 25. doi: 10.1038/nbt.2908. Csy4 and Cas9 nuclease expression plasmid pCAG-CFP
    pSQT817Cas9CAG Joung Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. Nat Biotechnol. 2014 Apr 25. doi: 10.1038/nbt.2908. wildtype Cas9 expression plasmid pCAG-CFP
    pL-CRISPR.EFS.GFPSpCas9 (Other), Sp sgRNA scaffold, EFS (Homo sapiens), P2A-eGFP (Synthetic)EFS, hU6 Ebert Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA. Coexpresses eGFP via P2A cleavage site. EFS Promoter driven pLKO.005
    pL-CRISPR.EFS.tRFPSpCas9 (Other), Sp sgRNA scaffold, EFS (Homo sapiens), P2A-tRFP (Synthetic) Ebert Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA. Coexpresses tagRFP via P2A cleavage site. EFS Promoter driven pLKO.005
    pLKO5d.EFS.SpCas9.P2A.BSDSpCas9 (Other), EFS (Homo sapiens), P2A-BSD (Synthetic)Blasticidin Ebert Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. Lentiviral Vector for SpCas9 Expression without sgRNA, Blasticidin resistance, EFS Promoter driven pLKO.005
    pL-CRISPR.SFFV.tRFPSpCas9 (Other), Sp sgRNA scaffold, SFFV (Synthetic), P2A-tRFP (Synthetic) Ebert Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA. Coexpresses tagRFP via P2A cleavage site. SFFV Promoter driven pLKO.005
    pL-CRISPR.SFFV.GFPSpCas9 (Other), Sp sgRNA scaffold, SFFV (Synthetic), P2A-eGFP (Synthetic) Ebert Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA. Coexpresses eGFP via P2A cleavage site. SFFV Promoter driven pLKO.005
    pL-CRISPR.EFS.PACSpCas9 (Other), Sp sgRNA scaffold, EFS (Homo sapiens), P2A-PAC (Synthetic)Puromycin Ebert Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA, Puromycin resistance, EFS Promoter driven pLKO.005
    pL-CRISPR.SFFV.PACSpCas9 (Other), Sp sgRNA scaffold, SFFV (Synthetic), P2A-PAC (Synthetic)Puromycin Ebert Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA, Puromycin resistance, SFFV Promoter driven pLKO.005
    pAdSh.PGK.Cas9Human codon-optimized S. pyogenes Cas9 (Synthetic)Neomycin (select with G418) Goncalves Adenoviral vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human cells. Sci Rep. 2014 May 29;4:5105. doi: 10.1038/srep05105. Expresses human codon-optimized S. pyogenes Cas9 from the human PGK-1 gene promoter pBR322-based pAdEasy backbone
    pLKO5d.EFS.SpCas9.P2A.PACSpCas9 (Other), EFS (Homo sapiens), P2A-PAC (Synthetic)Puromycin Ebert Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. Lentiviral Vector for SpCas9 Expression without sgRNA, Puromycin resistance, EFS Promoter driven pLKO.005
    Puro-Cas9 donorhSpCas9 (Other)Puromycin Huangfu An iCRISPR Platform for Rapid, Multiplexable, and Inducible Genome Editing in Human Pluripotent Stem Cells. Cell Stem Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 10.1016/j.stem.2014.05.018. Donor vector for genomic targeting of a Tetracycline-inducible Cas9 cassette to the human AAVS1/PPP1R12C locus N/A
    pX330A-1x2humanized S. pyogenes Cas9 nuclease (Other)CBh Yamamoto Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400. Expresses Cas9 nuclease and gRNA pUC ori vector
    piCRg EntryhSpCas9 Huangfu An iCRISPR Platform for Rapid, Multiplexable, and Inducible Genome Editing in Human Pluripotent Stem Cells. Cell Stem Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 10.1016/j.stem.2014.05.018. Cas9/gRNA Entry expression plasmid Low copy
    pCAG-T3-hCASeGFP-pACAS9 (Synthetic)CAG Fujii Efficient generation of large-scale genome-modified mice using gRNA and CAS9 endonuclease. Nucleic Acids Res. 2013 Aug 30. Expresses CAS9-eGFP fusion protein in mammalian cells. pCAGGS
    Eef1a-1955/-1>nls::Cas9::nlsnls::Cas9::nls (Other)Ciinte.Eef1a (EF1alpha) -1955/-1 Christiaen Tissue-specific genome editing in Ciona embryos by CRISPR/Cas9. Development. 2014 Nov;141(21):4115-20. doi: 10.1242/dev.114488. Eef1a (EF1alpha) promoter driving nls::Cas9::nls pCESAx
    Mesp-1916/-1>nls::Cas9::nlsnls::Cas9::nls (Other)Ciinte.Mesp -1916/-1 Christiaen Tissue-specific genome editing in Ciona embryos by CRISPR/Cas9. Development. 2014 Nov;141(21):4115-20. doi: 10.1242/dev.114488. Mesp driver driving nls::Cas9::nls pCESAx
    Sox1/2/3-2373/+3>nls::Cas9::nlsnls::Cas9::nls (Other)Ciinte.Sox1/2/3 (SoxB1) -2373/+3 Christiaen Tissue-specific genome editing in Ciona embryos by CRISPR/Cas9. Development. 2014 Nov;141(21):4115-20. doi: 10.1242/dev.114488. Sox1/2/3 (SoxB1) driver driving nls::Cas9::nls pCESAx
    pHL-EF1a-SphcCas9-iP-ACRISPR Cas9 (Other)Puromycin Hotta Precise Correction of the Dystrophin Gene in Duchenne Muscular Dystrophy Patient Induced Pluripotent Stem Cells by TALEN and CRISPR-Cas9. Stem Cell Reports. 2014 Nov 25. pii: S2213-6711(14)00335-X. doi: 10.1016/j.stemcr.2014.10.013. Expresses human codon-optimized Cas9 (derived from Streptococcus pyogenes) and pruomycin resistance gene. pHL-EF1a-GW-iP-A
    pSECCCas9 (Other), CreEFS, EFS (after Cas9-2A) Jacks Rapid modelling of cooperating genetic events in cancer through somatic genome editing. Nature. 2014 Oct 22. doi: 10.1038/nature13906. 3rd generation vector. Expresses a sgRNA of interest, Cas9 and Cre pLL3.3
    PX551SpCas9 (Synthetic)pMecp2 Zhang In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9. Nat Biotechnol. 2014 Oct 19. doi: 10.1038/nbt.3055. pAAV-pMecp2-SpCas9-spA (AAV-SpCas9). AAV plasmid expressing Cas9 in neurons under control of truncated mecp2 promoter. pAAV
    LSL-Cas9-Rosa26TVCas9 (Synthetic), EGFPCAG, CAGNeomycin (select with G418) ; DTA Zhang CRISPR-Cas9 Knockin Mice for Genome Editing and Cancer Modeling. Cell. 2014 Sep 24. pii: S0092-8674(14)01163-5. doi: 10.1016/j.cell.2014.09.014. Cre-dependent Cas9 expression plasmid and targeting vector for the mouse Rosa26 locus. Cas9 expression is Cre-dependent and the targeting vector has positive (Neo) and negative (DTA) selection. Ai9
    pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNAhSaCas9 (Other), Chimeric guide for SaCas9 (Other) Zhang In vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015 Apr 1. doi: 10.1038/nature14299. A single vector AAV-Cas9 system containing Cas9 from Staphylococcus aureus (SaCas9) and its sgRNA. pAAV
    pX600-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpAhSaCas9 (Other) Zhang In vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015 Apr 1. doi: 10.1038/nature14299. A human codon-optimized Cas9 from Staphylococcus aureus (SaCas9). pAAV
    pX602-AAV-TBG::NLS-SaCas9-NLS-HA-OLLAS-bGHpA;U6::BsaI-sgRNAhSaCas9 (Other), Chimeric guide for SaCas9 (Other) Zhang In vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015 Apr 1. doi: 10.1038/nature14299. A single vector AAV-Cas9 system containing Cas9 from Staphylococcus aureus (SaCas9) and its sgRNA. pAAV
    pMuLE ENTR CMV-hCas9 R4-R3human codon optimized Cas9 (Synthetic)CMV Frew A versatile modular vector system for rapid combinatorial mammalian genetics. J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743. MuLE (Multiple Lentiviral Expression) Entry vector containing CMV promoter and human codon optimized Cas9 module Compatible with MultiSite Gateway cloning N/A
    pMuLE ENTR SV40-hCas9 L3-L2human codon optimized Cas9 (Synthetic)SV40 Frew A versatile modular vector system for rapid combinatorial mammalian genetics. J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743. MuLE (Multiple Lentiviral Expression) Entry vector containing SV40 promoter and human codon optimized Cas9 module. Compatible with MultiSite Gateway cloning N/A
    pMuLE ENTR SV40-hCas9 L5-L2human codon optimized Cas9 (Synthetic)SV40 Frew A versatile modular vector system for rapid combinatorial mammalian genetics. J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743. MuLE (Multiple Lentiviral Expression) Entry vector containing SV40 promoter and human codon optimized Cas9 module. Compatible with MultiSite Gateway cloning N/A
    c3GIC9humanized S. Pyogenes Cas9 (Other)TRE3G Dow Inducible in vivo genome editing with CRISPR-Cas9. Nat Biotechnol. 2015 Apr;33(4):390-4. doi: 10.1038/nbt.3155. Epub 2015 Feb 18. col1a1 targeting vector for inducible [TRE3G]-GFP-IRES-Cas9 expression. Contains NsiI cloning site for U6-sgRNA cassettes col1a1 Flp-in targeting construct
    ptreCas9-mKate2ps-T1gRNACas9–mKate2ps (Synthetic) Bleris CRISPR-based self-cleaving mechanism for controllable gene delivery in human cells. Nucleic Acids Res. 2015 Jan 30;43(2):1297-303. doi: 10.1093/nar/gku1326. Epub 2014 Dec 18. Inducible; gRNA seq is ATGAGAATCAAGGCGGTCGA pTag-CFP
    pCas9–mKate2ps–EgRNACas9–mKate2ps (Synthetic) Bleris CRISPR-based self-cleaving mechanism for controllable gene delivery in human cells. Nucleic Acids Res. 2015 Jan 30;43(2):1297-303. doi: 10.1093/nar/gku1326. Epub 2014 Dec 18. Empty; no U6 gRNA cassette pTag-CFP
    pCas9–mKate2ps–T1gRNACas9–mKate2ps (Synthetic) Bleris CRISPR-based self-cleaving mechanism for controllable gene delivery in human cells. Nucleic Acids Res. 2015 Jan 30;43(2):1297-303. doi: 10.1093/nar/gku1326. Epub 2014 Dec 18. t1 gRNA (ATGAGAATCAAGGCGGTCGA) pTag-CFP
    PX851SpCas9 (aa 2-535)CBh Zhang A split-Cas9 architecture for inducible genome editing and transcription modulation. Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149. N-term SpCas9 piece of inducible split-4 (Cas9(N)-FRB-NES split-4). PX330
    PX852SpCas9 (aa536-1368)CBh Zhang A split-Cas9 architecture for inducible genome editing and transcription modulation. Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149. C-term SpCas9 piece of inducible split-4 (Cas9(C)-FKBP split-4). PX330
    PX853SpCas9 (aa 2-573)CBh Zhang A split-Cas9 architecture for inducible genome editing and transcription modulation. Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149. N-term SpCas9 piece of inducible split-5 (Cas9(N)-FRB-NES split-5). PX330
    PX854SpCas9 (aa573-1368)CBh Zhang A split-Cas9 architecture for inducible genome editing and transcription modulation. Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149. C-term SpCas9 piece of inducible split-5 (Cas9(C)-FKBP split-5). PX330
    pLSC-5SpCas9(574-1368), SpCas9(2-573)EFS, EFSPuromycin Zhang A split-Cas9 architecture for inducible genome editing and transcription modulation. Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149. Lenti vector for expression of inducible split Cas9 lentiCRISPR
    pSpCas9(BB)-2A-Puro (PX459) V2.0hSpCas9-2A-Puro V2.0 (Synthetic)CbhPuromycin Zhang Genome engineering using the CRISPR-Cas9 system. Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24. Cas9 from S. pyogenes with 2A-Puro, and cloning backbone for sgRNA (V2.0) PX459
    lentiCas9-EGFPCas9 (Synthetic), EGFP (Other)EFS-NS, EFS-NS Zhang Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis. Cell. 2015 Mar 12;160(6):1246-60. doi: 10.1016/j.cell.2015.02.038. Epub 2015 Mar 5. Expresses human codon-optimized S. pyogenes Cas9 protein and EGFP from EFS promoter. 3rd generation lentiviral backbone. pFUGW
    pX330S-2-PITChhumanized S. pyogenes Cas9 (Other)CBh Yamamoto MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems. Nat Protoc. 2016 Jan;11(1):118-33. doi: 10.1038/nprot.2015.140. Epub 2015 Dec 17. Expresses Cas9 nuclease and the PITCh-gRNA pUC ori vector
    pX330A-FBL/PITChhumanized S. pyogenes Cas9 (Other)CBh Yamamoto MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems. Nat Protoc. 2016 Jan;11(1):118-33. doi: 10.1038/nprot.2015.140. Epub 2015 Dec 17. Expresses Cas9 nuclease, FBL-specific gRNA, and the PITCh-gRNA pUC ori vector
    PX458_CEBPB_1gRNAU6 Mendenhall CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res. 2015 Sep 9. Expresses gRNA against human CEBPB along with Cas9 with 2A GFP px548
    PX458_CEBPB_2gRNAU6 Mendenhall CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res. 2015 Sep 9. Expresses gRNA against human CEBPB along with Cas9 with 2A GFP px548
    PX458_RAD21_1gRNA Mendenhall CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res. 2015 Sep 9. Expresses gRNA against human RAD21 along with Cas9 with 2A GFP px548
    Adeno Cas9U6 and CBh Ventura In vivo engineering of oncogenic chromosomal rearrangements with the CRISPR/Cas9 system. Nature. 2014 Dec 18;516(7531):423-7. doi: 10.1038/nature13902. Epub 2014 Oct 22. Adenoviral vector for the expression of the Cas9 pacAd5
  • Tag / Fusion Protein
    • FLAG
  • pX333CBh; U6 Ventura In vivo engineering of oncogenic chromosomal rearrangements with the CRISPR/Cas9 system. Nature. 2014 Dec 18;516(7531):423-7. doi: 10.1038/nature13902. Epub 2014 Oct 22. Vector for tandem expression of two sgRNAs from two independent U6 promoters. Cas9 is expressed by Cbh promoter. pX333
  • Tag / Fusion Protein
    • 3XFLAG-Cas9 (N terminal on backbone)
  • pKMD106e - intein-Cas9(S219)mammalian codon-optimized S. pyogenes intein-Cas9(S219) - NLS - 3x FLAG (Other)CMV Liu Small molecule-triggered Cas9 protein with improved genome-editing specificity. Nat Chem Biol. 2015 May;11(5):316-8. doi: 10.1038/nchembio.1793. Epub 2015 Apr 6. Expresses intein-Cas9(S219) in mammalian cells pJDS246
    pKMD106h - intein-Cas9(C574)mammalian codon-optimized S. pyogenes intein-Cas9(C574) - NLS - 3x FLAG (Other)CMV Liu Small molecule-triggered Cas9 protein with improved genome-editing specificity. Nat Chem Biol. 2015 May;11(5):316-8. doi: 10.1038/nchembio.1793. Epub 2015 Apr 6. Expresses intein-Cas9(C574) in mammalian cells pJDS246
    pKMD111e - intein-Cas9(S219-G521R)mammalian codon-optimized S. pyogenes intein-Cas9(S219-G521R) - NLS - 3x FLAG (Other)CMV Liu Small molecule-triggered Cas9 protein with improved genome-editing specificity. Nat Chem Biol. 2015 May;11(5):316-8. doi: 10.1038/nchembio.1793. Epub 2015 Apr 6. Expresses intein-Cas9(S219-G521R) in mammalian cells pJDS246
    pU6-sgRosa26-1_CBh-Cas9-T2A-BFPCas9 (Other), sgRNA targeting ROSA26-1CBh, U6 Kuehn Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. Expression vector for a sgRNA against the mouse Rosa26 locus and Cas9 linked to BFP via a T2A peptide pU6-(BbsI)_CBh-Cas9-T2A-BFP
    pU6-(BbsI)_CBh-Cas9-T2A-mCherry-H1-(BamHI)Cas9 (Other), hU6 promoter; BbsI sites for sgRNA, H1 promoter; BamHI site for shRNACBh, U6, H1 Kuehn Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. Expression vector for sgRNAs cloned into the BbsI sites, shRNAs into BamHI & AflII and for Expression of Cas9 linked to mCherry via T2A pU6-(BbsI)_CBh-Cas9-T2A-BFP
    pU6-(BbsI)_CBh-Cas9-T2A-BFP-P2A-Ad4E1BCas9 (Other), sgRNA cassetteCBh, U6 Kuehn Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. Expression vector for sgRNA and for Expression of Cas9 linked to BFP via T2A linked to Ad4 E1B via P2A pU6-(BbsI)_CBh-Cas9-T2A-BFP
    pU6-sgRosa26-1_CBh-Cas9-T2A-BFP-P2A-Ad4E1BCas9 (Other), sgRNA targeting ROSA26-1CBh, U6 Kuehn Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. Expression vector for a sgRNA against the mouse Rosa26 locus and Cas9 linked via T2A to BFP linked to the Ad4 E1B55K gene via P2A pU6-(BbsI)_CBh-Cas9-T2A-BFP-P2A-Ad4E1B
    pU6-(BbsI)_CBh-Cas9-T2A-BFP-P2A-Ad4E4orf6Cas9 (Other), sgRNA cassetteCBh, U6 Kuehn Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. Expression vector for sgRNA and Cas9 linked via T2A to BFP linked to the Ad4 E4orf6 gene via P2A pU6-(BbsI)_CBh-Cas9-T2A-BFP
    pU6-(BbsI)_CBh-Cas9-T2A-mcherry-P2A-Ad4E1BCas9 (Other), sgRNA cassetteCBh, U6 Kuehn Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. Expression vector for sgRNA and for Expression of Cas9 linked via T2A to mCherry and to Ad4 E1B55K via P2A pU6-(BbsI)_CBh-Cas9-T2A-mCherry
    pU6-(BbsI)_CBh-Cas9-T2A-mcherry-P2A-Ad4E4orf6Cas9 (Other), sgRNA cassetteCBh, U6 Kuehn Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. Expression vector for sgRNA and for Expression of Cas9 linked via T2A to mCherry linked to the Ad4 E4orf6 gene via P2A pU6-(BbsI)_CBh-Cas9-T2A-mCherry
    PX458_GABPA_1gRNA Mendenhall CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res. 2015 Sep 9. Expresses gRNA against human GABPA along with Cas9 with 2A GFP px548
    PX458_GABPA_2gRNA Mendenhall CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res. 2015 Sep 9. Expresses gRNA against human GABPA along with Cas9 with 2A GFP px548
    pU6-(BbsI)_CBh-Cas9-T2A-BFPCas9 (Other), sgRNA cassetteCBh, U6 Kuehn Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. Expression vector for sgRNAs cloned into the BbsI sites and for expression of Cas9 linked to BFP via a T2A peptide pX330
    pU6-(BbsI)_CBh-Cas9-T2A-mCherryCas9 (Other), sgRNA cassetteCBh, U6 Kuehn Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. Expression vector for sgRNAs cloned into the BbsI sites and for expression of Cas9 linked to mCherry via a T2A peptide pX330
    pMCS-rybozyme-IRES-CAS9CAS9 (Synthetic), HDV ribozyme (Other) Fujii Development of a mono-promoter-driven CRISPR/Cas9 system in mammalian cells. Sci Rep. 2015 Dec 16;5:18341. doi: 10.1038/srep18341. Plasmid encoding multiple cloning site, rybozyme and IRES-CAS9. pCAGGS
    PX458_ATF1_1gRNA Mendenhall CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res. 2015 Sep 9. Expresses gRNA against human ATF1 along with Cas9 with 2A GFP px548
    pSaCas9_GFPSaCas9-2A-GFP (Synthetic)CAG Musunuru Staphylococcus aureus Cas9 (unpublished) Co-expression of human codon-optimized Staphylococcus aureus Cas9 nuclease and GFP, plasmid optimized for expression in human pluripotent stem cells and other mammalian cells pCAG
    PX458_CREB1_1gRNA Mendenhall CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res. 2015 Sep 9. Expresses gRNA against human CREB1 along with Cas9 with 2A GFP px548
    PX458_CREB1_2gRNA Mendenhall CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res. 2015 Sep 9. Expresses gRNA against human CREB1 along with Cas9 with 2A GFP px548
    pCas9–mKate2ps–CgRNACas9–mKate2ps Bleris CRISPR-based self-cleaving mechanism for controllable gene delivery in human cells. Nucleic Acids Res. 2015 Jan 30;43(2):1297-303. doi: 10.1093/nar/gku1326. Epub 2014 Dec 18. Control gRNA (GTCAAGGCACTCTTGCCTA) pTag-CFP
    MSCV_Cas9_puroCas9 (Synthetic)MSSV_LTRPuromycin Vakoc Discovery of cancer drug targets by CRISPR-Cas9 screening of protein domains. Nat Biotechnol. 2015 May 11. doi: 10.1038/nbt.3235. Retroviral introduction of Cas9 into mammalian cell line. MSCV_puro
    MSP469mammalian codon-optimized Streptococcus pyogenes Cas9 (D1135V/R1335Q/T1337R)-NLS-3XFlag (Other)CMV & T7 Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Human expression vector for SpCas9 VQR variant: CMV-T7-humanSpCas9(D1135V/R1335Q/T1337R)-NLS-3xFLAG (VQR variant) JDS246
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)
  • MSP680mammalian codon-optimized Streptococcus pyogenes Cas9 (D1135E/R1335Q/T1337R)-NLS-3XFlag (Other)CMV & T7 Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Human expression vector for SpCas9 EQR variant: CMV-T7-humanSpCas9(D1135E/R1335Q/T1337R)-NLS-3xFLAG (EQR variant) JDS246
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)
  • MSP1101mammalian codon-optimized Streptococcus pyogenes Cas9 (D1135V/G1218R/R1335E/T1337R)-NLS-3XFlag (Other)CMV & T7 Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Human expression vector for SpCas9 VRER variant: CMV-T7-humanSpCas9(D1135V/G1218R/R1335E/T1337R)-NLS-3xFLAG (VRER variant) JDS246
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)
  • MSP977mammalian codon-optimized Streptococcus pyogenes Cas9 (D1135E)-NLS-3XFlag (Other)CMV & T7 Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Human expression vector for SpCas9 D1135E variant: CMV-T7-humanSpCas9(D1135E)-NLS-3xFLAG JDS246
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)
  • MSP1594mammalian codon-optimized Streptococcus thermophilus1 Cas9-NLS (Other)CAG Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Human expression vector for S. thermophilus 1 Cas9: CAG-humanSt1Cas9-NLS pCAG-CFP
  • Tag / Fusion Protein
    • NLS (C terminal on insert)
  • BPK2139mammalian codon-optimized Staphylococcus aureus Cas9-NLS-3xFlag (Other)CAG Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Human expression vector for S .aureus Cas9: CAG-humanSaCas9-NLS-3xFLAG pCAG-CFP
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)
  • BPK1520SpCas9 gRNA backbone, without spacer sequence (Synthetic)U6 Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Human expression plasmid for SpCas9 sgRNA (need to clone in spacer into BsmBI sites): U6-BsmBIcassette-Sp-sgRNA MLM3636
    BPK2301St1Cas9 gRNA backbone, without spacer sequence (Synthetic)U6 Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Human expression plasmid for S. thermophilus 1 Cas9 sgRNA (need to clone in spacer into BsmBI sites): U6-BsmBIcassette-St1-sgRNA MLM3636
    VVT1SaCas9 gRNA backbone, without spacer sequence (Synthetic)U6 Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. The Joung Lab recommends using BPK2660 (Addgene plasmid 70709) instead of VVT1 as guide RNAs cloned into BPK2660 are more effective than this plasmid. Human expression plasmid for S. aureus Cas9 sgRNA MLM3636
    pBGKCas9 Ohtsuka Mouse Genome Editing Using the CRISPR/Cas System. Curr Protoc Hum Genet. 2014 Oct 1;83:15.7.1-15.7.27. doi: 10.1002/0471142905.hg1507s83. Vector for Cas9 mRNA synthesis pcDNA3.1
    pX330.iGFP1iGFP Xue A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome Biol. 2015 May 28;16(1):111. sgRNA for iGFP pX330
    pX330.iGFP2iGFP Xue A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome Biol. 2015 May 28;16(1):111. sgRNA for iGFP pX330
    pX330.iGFP3iGFP Xue A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome Biol. 2015 May 28;16(1):111. sgRNA for iGFP pX330
    pX330.iGFP5iGFP Xue A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome Biol. 2015 May 28;16(1):111. sgRNA for iGFP pX330
    pX330.LoxPOLSL-Tomato Xue A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome Biol. 2015 May 28;16(1):111. sgRNA for LSL pX330
    pX330.LoxPLoxP sgRNA Xue A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome Biol. 2015 May 28;16(1):111. sgRNA for LSL pX330
    pX330.Pten.aPten deletion (Mus musculus) Xue A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome Biol. 2015 May 28;16(1):111. sgRNA for Pten deletion pX330 Pten 2310035O07Rik, A130070J02Rik, AI463227, B430203M17Rik, MMAC1, PTENbeta, TEP1
    pX330.Pten.bPten deletion (Mus musculus) Xue A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome Biol. 2015 May 28;16(1):111. sgRNA for Pten deletion pX330 Pten 2310035O07Rik, A130070J02Rik, AI463227, B430203M17Rik, MMAC1, PTENbeta, TEP1
    pCAS9-mCherry-Frame +0gRNA frame +0 Hornung CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. CRISPaint frame selector +0 pCAS9-mCherry
    pCAS9-mCherry-Frame +1gRNA frame +1 Hornung CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. CRISPaint frame selector +1 pCAS9-mCherry
    pCAS9-mCherry-Frame +2gRNA frame +2 Hornung CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. CRISPaint frame selector +2 pCAS9-mCherry
    pCAS9-mCherry-TUBBgRNA TUBB Hornung CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. CRISPaint target selector TUBB pCAS9-mCherry
    pCAS9-mCherry-ACTG1gRNA ACTG1 (Homo sapiens) Hornung CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. CRISPaint target selector ACTG1 pCAS9-mCherry
    pCAS9-mCherry-HIST1H4CgRNA HIST1H4C (Homo sapiens) Hornung CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. CRISPaint target selector HIST1H4C pCAS9-mCherry
    pKLV2-EF1a-BsdCas9-WEF1a-Cas9 cassette, WPRE (Homo sapiens)EF1a Yusa A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia. Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 10.1016/j.celrep.2016.09.079. Lentiviral vector expressing Cas9 fused with the Blasticidin resistant gene at the N-terminus pKLV2 lentiviral vector
    pKLV2-U6gRNA5(Empty)-PGKGFP2ABFP-WU6gRNA cassette, PGKGFP2ABFP cassette, WPRE (Homo sapiens) Yusa A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia. Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 10.1016/j.celrep.2016.09.079. Cas9 activity reporter (control) with GFP and BFP pKLV2 lentiviral vector
    pRosa26-EF1a-hCas9IRESneoEF1a-hCas9IRESneopA (Synthetic) Yusa A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia. Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 10.1016/j.celrep.2016.09.079. Mouse Rosa26 targeting vector carrying the EF1a-hCas9-IRES-neo cassette pBluescriptII
    Mouse genome-wide lentiviral CRISPR gRNA library version 2U6gRNA cassette, PGKpuro2ABFP cassette, WPRE (Homo sapiens), Mouse CRISPR guide RNA library v2 (Synthetic) Yusa A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia. Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 10.1016/j.celrep.2016.09.079. Mouse genome-wide CRISPR guide RNA library V2. (pKLV2-U6gRNA5(MouseV2)-PGKpuro2ABFP-W) pKLV2 lentiviral vector
    Human genome-wide lentiviral CRISPR gRNA library version 1U6gRNA cassette, PGKpuro2ABFP cassette, WPRE (Homo sapiens), Human CRISPR guide RNA library v2 (Synthetic) Yusa A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia. Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 10.1016/j.celrep.2016.09.079. Human genome-wide CRISPR guide RNA library V1 (pKLV2-U6gRNA5(HumanV1)-PGKpuro2ABFP-W) pKLV2 lentiviral vector
    phumanCas9 (GB0575)Cas9 coding region (human codon optimised) (Other) Orzaez GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. Plant Physiol. 2013 Jul;162(3):1618-31. Provides the human codon optimized CDS of Cas9 protein as a level 0 GoldenBraid part pUPD
    PX377 Corynebacter diphtheriae Cas9CdCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Corynebacter diphtheriae Cas9 pDREAM
    PX378 Sutterella wadsworthensis Cas9SwCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Sutterella wadsworthensis Cas9 pDREAM
    PX379 Legionella pneumophila str. Paris Cas9LpCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Legionella pneumophila str. Paris Cas9 pDREAM
    PX380 Treponema denticola Cas9TdCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Treponema denticola Cas9 pDREAM
    PX381 Filifactor alocis Cas9FaCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Filifactor alocis Cas9 pDREAM
    PX382 Staphylococcus pseudintermedius Cas9SpsCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Staphylococcus pseudintermedius Cas9 pDREAM
    PX383 Lactobacillus johnsonii Cas9LjCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Lactobacillus johnsonii Cas9 pDREAM
    PX385 Streptococcus pasteurianus Cas9SpaCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Streptococcus pasteurianus Cas9 pDREAM
    PX386 Lactobacillus farciminis Cas9LfCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Lactobacillus farciminis Cas9 pDREAM
    PX387 Mycoplasma mobile Cas9MmCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Mycoplasma mobile Cas9 pDREAM
    PX388 Bacteroides coprophilus Cas9BcCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Bacteroides coprophilus Cas9 pDREAM
    PX389 Fluviicola taffensis Cas9FtCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Fluviicola taffensis Cas9 pDREAM
    PX390 Flavobacterium columnare Cas9FcCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Flavobacterium columnare Cas9 pDREAM
    PX391 Sphaerochaeta globus str. Buddy Cas9SgCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Sphaerochaeta globus str. Buddy Cas9 pDREAM
    PX392 Azospirillum B510 Cas9AzoCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Azospirillum B510 Cas9 pDREAM
    PX393 Gluconacetobacter diazotrophicus Cas9GdCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Gluconacetobacter diazotrophicus Cas9 pDREAM
    PX394 Neisseria cinerea Cas9NcCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Neisseria cinerea Cas9 pDREAM
    PX395 Roseburia intestinalis Cas9RiCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Roseburia intestinalis Cas9 pDREAM
    PX396 Parvibaculum lavamentivorans Cas9PlCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Parvibaculum lavamentivorans Cas9 pDREAM
    PX398 Nitratifractor salsuginis str DSM 16511 Cas9NsCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Nitratifractor salsuginis str DSM 16511 Cas9 pDREAM
    PX399 Mycoplasma gallisepticum str. F Cas9MgCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Mycoplasma gallisepticum str. F Cas9 pDREAM
    PX400 Campylobacter lari CF89-12 Cas9ClCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Campylobacter lari CF89-12 Cas9 pDREAM
    PX403 Streptococcus thermophilus LMD-9 CRISPR 3 Cas9St3Cas9 (Synthetic)CBh Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Streptococcus thermophilus LMD-9 CRISPR 3 Cas9 Unknown
    PX404 Campylobacter jejuni Cas9CjCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Campylobacter jejuni Cas9 Unknown
    pKLV2-EF1a-Cas9Bsd-WEF1a-Cas9 cassette, WPRE (Homo sapiens)EF1a Yusa A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia. Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 10.1016/j.celrep.2016.09.079. Lentiviral vector expressing Cas9 fused with the Blasticidin resistant gene at the C-terminus pKLV2 lentiviral vector
    LIR_TRE_CAG_mKateII-Puro-STOP_mVenus-Cas9-PA-RIRpuromycin (Synthetic), hCas9 (Synthetic)CAG, CAGPuromycin Elverløv-Jakobsen Cas9/CRISPR pig transposon plasmid_LIR_TRE_CAG_mKateII-Puro-STOP_mVenus-Cas9-PA-RIR (unpublished) A Sleeping beauty transposon with conditional expressed hCas9. A red flourescent gene linked to puromycin can be removed in the prescence of Flp recombinase allowing Cas9 expression Amp
    px330_P53 exon 8 gRNAP53 exon 8 gRNA (Synthetic)U6 Elverløv-Jakobsen Cas9/CRISPR pig transposon plasmid_LIR_TRE_CAG_mKateII-Puro-STOP_mVenus-Cas9-PA-RIR (unpublished) px330 with gRNA towards P53 exon 8. Cas9 is expressed from a CAG promoter. amp
    px330_P53 exon 7 gRNAP53 exon 7 gRNA (Synthetic)U6 Elverløv-Jakobsen Cas9/CRISPR pig transposon plasmid_LIR_TRE_CAG_mKateII-Puro-STOP_mVenus-Cas9-PA-RIR (unpublished) px330 with gRNA towards P53 exon 7. Cas9 is expressed from a CAG promoter. amp
    px330_SMAD exon 9 gRNASMAD exon 9 gRNA (Synthetic)U6 Elverløv-Jakobsen Cas9/CRISPR pig transposon plasmid_LIR_TRE_CAG_mKateII-Puro-STOP_mVenus-Cas9-PA-RIR (unpublished) px330 with gRNA towards SMAD exon 9. Cas9 is expressed from a CAG promoter. amp
    CAS9PBKSCas9 (Other)CbAkt Bennett Genome editing using FACS enrichment of nuclease-expressing cells and indel detection by amplicon analysis. Nat Protoc. 2017 Mar;12(3):581-603. doi: 10.1038/nprot.2016.165. Epub 2017 Feb 16. CbAkt promoter driven Cas9-2A-GFP (from px458) pBKS
    Cas9-2A-CreCas9 and Cre recombinase (Homo sapiens)CAG Zhang Engineering Human Stem Cell Lines with Inducible Gene Knockout using CRISPR/Cas9. Cell Stem Cell. 2015 Jul 1. pii: S1934-5909(15)00261-1. doi: 10.1016/j.stem.2015.06.001. co-expressing Cas9 and Cre recombinase Cas9-2A-GFP
    PX405 Neisseria meningitidis Cas9NmCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Neisseria meningitidis Cas9 pDREAM
    PX406 Pasteurella multocida Cas9PmCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Pasteurella multocida Cas9 pDREAM
    PX408 Francisella tularensis subsp. novicida Cas9FtCas9 (Synthetic)CMV Zhang Feng Zhang lab bacterial Cas9 orthologs (unpublished) Francisella tularensis subsp. novicida Cas9 pDREAM
    pCSDest2-SpCas9-NLS-3XHA-NLS-Zif268Zif268 (Mus musculus) Wolfe DNA-binding-domain fusions enhance the targeting range and precision of Cas9. Nat Methods. 2015 Oct 19. doi: 10.1038/nmeth.3624. Expresses wild type SpCas9 fused to Zif268 in mammalian cells pCSDest2-SpCas9-NLS-3XHA-NLS Egr1 A530045N19Rik, ETR103, Egr-1, Krox-1, Krox-24, Krox24, NGF1-A, NGFI-A, NGFIA, TIS8, Zenk, Zfp-6, Zif268, egr
    pCSDest2-SpCas9-NLS-3XHA-NLSSpCas9 (Other) Wolfe DNA-binding-domain fusions enhance the targeting range and precision of Cas9. Nat Methods. 2015 Oct 19. doi: 10.1038/nmeth.3624. Expresses wild type SpCas9 in mammalian cells pCSDest2
    pCSDest2-SpCas9-WT-NLS-3XHA-NLS-ZFP_TS2TS2 ZFP (Synthetic) Wolfe DNA-binding-domain fusions enhance the targeting range and precision of Cas9. Nat Methods. 2015 Oct 19. doi: 10.1038/nmeth.3624. Expresses wild type SpCas9 fused to ZFP-TS2 in mammalian cells pCSDest2-SpCas9-NLS-3XHA-NLS
    pCSDest2-SpCas9-WT-NLS-3XHA-NLS-ZFP_TS3TS3 ZFP (Synthetic) Wolfe DNA-binding-domain fusions enhance the targeting range and precision of Cas9. Nat Methods. 2015 Oct 19. doi: 10.1038/nmeth.3624. Expresses wild type SpCas9 fused to ZFP-TS3 in mammalian cells pCSDest2-SpCas9-NLS-3XHA-NLS
    pCSDest2-SpCas9-WT-NLS-3XHA-NLS-ZFP_TS4TS4 ZFP (Synthetic) Wolfe DNA-binding-domain fusions enhance the targeting range and precision of Cas9. Nat Methods. 2015 Oct 19. doi: 10.1038/nmeth.3624. Expresses wild type SpCas9 fused to ZFP-TS4 in mammalian cells pCSDest2-SpCas9-NLS-3XHA-NLS
    pCSDest2-SpCas9-MT3-NLS-3XHA-NLS-ZFP_TS3TS3 ZFP (Synthetic) Wolfe DNA-binding-domain fusions enhance the targeting range and precision of Cas9. Nat Methods. 2015 Oct 19. doi: 10.1038/nmeth.3624. Expresses R1335K mutant SpCas9 fused to ZFP-TS3 in mammalian cell pCSDest2-SpCas9-MT3-NLS-3XHA-NLS-Zif268
    FUCas9Cherryhumanised S.pyogenes cas9 (Other)hUbC Herold An inducible lentiviral guide RNA platform enables the identification of tumor-essential genes and tumor-promoting mutations in vivo. Cell Rep. 2015 Mar 3;10(8):1422-32. doi: 10.1016/j.celrep.2015.02.002. Epub 2015 Feb 26. Expresses Cas9 with fluorescent Cherry reporter FUGW
    lentiCas9-VenusCas9 (Synthetic), Venus Reporter (Synthetic)EFS-NS, P2AVenus Bauer BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis. Nature. 2015 Sep 16. doi: 10.1038/nature15521. Expresses human codon-optimized S. pyogenes Cas9 protein and Venus reporter from EFS promoter. Lentiviral backbone. pFUGW
    MSP1830mammalian codon-optimized KKH variant Staphylococcus aureus Cas9(E782K/N968K/R1015H)-NLS-3xFlag (Other)CAG Joung Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404. Human expression plasmid for SaCas9 KKH variant: CAG-humanSaCas9(E782K/N968K/R1015H)-NLS-3xFLAG pCAG-CFP
    spCas9-BlastR (pCBhCas9-BlastR)spCas9CBhBlasticidin Sherwood Cloning-free CRISPR. Stem Cell Reports. 2015 Oct 27. pii: S2213-6711(15)00284-2. doi: 10.1016/j.stemcr.2015.09.022. Expresses SpCas9 and Blasticidin resistance p2Tol2
    pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110)humanized S. pyogenes Cas9 fused to human Geminin (Homo sapiens)CBh Draetta Post-translational Regulation of Cas9 during G1 Enhances Homology-Directed Repair. Cell Rep. 2016 Feb 3. pii: S2211-1247(16)00040-1. doi: 10.1016/j.celrep.2016.01.019. A human codon-optimized SpCas9 fused to first 110 amino acids of human GEMININ and chimeric guide RNA expression plasmid pX330-U6-Chimeric_BB-CBh-hSpCas9 GMNN Gem, MGORS6
    eSpCas9(1.1)enhanced specificity Cas9 (1.1) (Other)CBh Zhang Rationally engineered Cas9 nucleases with improved specificity. Science. 2015 Dec 1. pii: aad5227. Expresses high specificity SpCas9. Px330-like plasmid. pX330-U6-Chimeric_BB-CBh-hSpCas9
    pCR1002T7pr_10xHis-MBP-TEV-SPyCas9 Corn Enhancing homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Nat Biotechnol. 2016 Jan 20. doi: 10.1038/nbt.3481. Express Streptococcus pyogenes Cas9 SPynCas9
    VP12mammalian codon-optimized Streptococcus pyogenes Cas9 HF1(N497A/R661A/Q695A/Q926A)-NLS-3xFlag (Other)CMV Joung High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016 Jan 6. doi: 10.1038/nature16526. Human expression plasmid for SpCas9-HF1 variant: CMV-T7-humanSpCas9-HF1(N497A, R661A, Q695A, Q926A)-NLS-3xFLAG JDS246
    MSP2135mammalian codon-optimized Streptococcus pyogenes Cas9 HF2(N497A/R661A/Q695A/Q926A/D1135E)-NLS-3xFlag (Other)CMV Joung High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016 Jan 6. doi: 10.1038/nature16526. Human expression plasmid for SpCas9-HF2 variant: CMV-T7-humanSpCas9-HF2(N497A, R661A, Q695A, Q926A, D1135E)-NLS-3xFLAG JDS246
    MSP2133mammalian codon-optimized Streptococcus pyogenes Cas9 HF4(Y450A/N497A/R661A/Q695A/Q926A)-NLS-3xFlag (Other)CMV Joung High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016 Jan 6. doi: 10.1038/nature16526. Human expression plasmid for SpCas9-HF4 variant: CMV-T7-humanSpCas9-HF4(Y450A, N497A, R661A, Q695A, Q926A)-NLS-3xFLAG JDS246
    MSP2440mammalian codon-optimized Streptococcus pyogenes Cas9 VQR-HF1(N497A/R661A/Q695A/Q926A/D1135V/R1335Q/T1337R)-NLS-3xFlag (Other)CMV Joung High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016 Jan 6. doi: 10.1038/nature16526. Human expression plasmid for SpCas9-VQR-HF1 variant: CMV-T7-humanSpCas9-VQR-HF1(N497A, R661A, Q695A, Q926A, D1135V, R1335Q, T1337R)-NLS-3xFLAG JDS246
    BPK2797mammalian codon-optimized Streptococcus pyogenes Cas9 VRQR(D1135V/G1218R/R1335Q/T1337R)-NLS-3xFlag (Other)CMV Joung High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016 Jan 6. doi: 10.1038/nature16526. Human expression plasmid for SpCas9-VRQR variant: CMV-T7-humanSpCas9-VRQR(D1135V, G1218R, R1335Q, T1337R)-NLS-3xFLAG JDS246
    MSP2443mammalian codon-optimized Streptococcus pyogenes Cas9 VRQR-HF1(N497A/R661A/Q695A/Q926A/D1135V/G1218R/R1335Q/T1337R)-NLS-3xFlag (Other)CMV Joung High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016 Jan 6. doi: 10.1038/nature16526. Human expression plasmid for SpCas9-VRQR-HF1 variant: CMV-T7-humanSpCas9-VRQR-HF1(N497A, R661A, Q695A, Q926A, D1135V, G1218R, R1335Q, T1337R)-NLS-3xFLAG JDS246
    pCas9-polyACas9-polyadenine (Other)T7 Mashimo ssODN-mediated knock-in with CRISPR-Cas for large genomic regions in zygotes. Nat Commun. 2016 Jan 20;7:10431. doi: 10.1038/ncomms10431. express hCas9 byT7 promoter with polyadenine tails pcDNA3.3-TOPO
    MSP2372mammalian codon-optimized Streptococcus pyogenes Cas9 (R661A/Q695A/Q926A)-NLS-3xFlag (Other)CMV Joung High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature. 2016 Jan 6. doi: 10.1038/nature16526. Human expression plasmid for SpCas9(R661A/Q695A/Q926A) variant: CMV-T7-humanSpCas9(R661A, Q695A, Q926A)-NLS-3xFLAG JDS246
    Lenti‐Cas9‐2A‐BlastCas9 (Synthetic)EFS-NSBlasticidin Moffat High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities. Cell. 2015 Dec 3;163(6):1515-26. doi: 10.1016/j.cell.2015.11.015. Epub 2015 Nov 25. Lentiviral vector expressing human codon-optimized S. pyogenes FLAG-tagged Cas9 and blasticidin resistance from EFS promoter lenti CRISPR pXPR_001
    pAAVS1-PDi-CRISPRnCas9 (Synthetic), rtTA (Synthetic)TRE, CAGPuromycin Conklin CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs. Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi: 10.1016/j.stem.2016.01.022. Epub 2016 Mar 10. Dox-inducible CRISPR nuclease (CRISPRn) knock in construct into the AAVS1 locus pAAVS1
    pAWp30EFSp-Cas9-P2A-Zeo Lu Multiplexed barcoded CRISPR-Cas9 screening enabled by CombiGEM. Proc Natl Acad Sci U S A. 2016 Feb 10. pii: 201517883. pFUGW-EFSp-Cas9-P2A-Zeo pFUGW
    pBLO1811_Cas9_noNLS_humanCas9 (Homo sapiens) Savage Profiling of engineering hotspots identifies an allosteric CRISPR-Cas9 switch. Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528. Human codon optimized plasmid to express Cas9 t2a mCherry w/o NLS and a sgRNA px330
    pBLO1806_Cas9_2xNLS_humanCas9 (Homo sapiens) Savage Profiling of engineering hotspots identifies an allosteric CRISPR-Cas9 switch. Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528. Human codon optimized plasmid to express Cas9 t2a mCherry w/2xNLS and a sgRNA px330
    pLentiCRISPR-EGFP Bornhauser Activation of concurrent apoptosis and necroptosis by SMAC mimetics for the treatment of refractory and relapsed ALL. Sci Transl Med. 2016 May 18;8(339):339ra70. doi: 10.1126/scitranslmed.aad2986. Derived from LentiCRISPR v1 but expresses EGFP instead of puromycin resistance lentiCRISPRv1
  • Tag / Fusion Protein
    • EGFP
  • pLentiCRISPR-tagBFP Bornhauser Activation of concurrent apoptosis and necroptosis by SMAC mimetics for the treatment of refractory and relapsed ALL. Sci Transl Med. 2016 May 18;8(339):339ra70. doi: 10.1126/scitranslmed.aad2986. Derived from LentiCRISPR v1 but expresses tagBFP instead of puromycin resistance lentiCRISPRv1
  • Tag / Fusion Protein
    • tagBFP
  • pLentiCRISPR-mCherry Bornhauser Activation of concurrent apoptosis and necroptosis by SMAC mimetics for the treatment of refractory and relapsed ALL. Sci Transl Med. 2016 May 18;8(339):339ra70. doi: 10.1126/scitranslmed.aad2986. Derived from LentiCRISPR v1 but expresses mCherry instead of puromycin resistance. 3rd generation. lentiCRISPRv1
  • Tag / Fusion Protein
    • mCherry
  • pLentiCRISPR-RFP657 Bornhauser Activation of concurrent apoptosis and necroptosis by SMAC mimetics for the treatment of refractory and relapsed ALL. Sci Transl Med. 2016 May 18;8(339):339ra70. doi: 10.1126/scitranslmed.aad2986. Derived from LentiCRISPR v1 but expresses RFP657 instead of puromycin resistance lentiCRISPRv1
  • Tag / Fusion Protein
    • RFP657
  • FUG-T2A-Cas9Cas9Ubiquitin Luikart A Retroviral CRISPR-Cas9 System for Cellular Autism-Associated Phenotype Discovery in Developing Neurons. Sci Rep. 2016 May 10;6:25611. doi: 10.1038/srep25611. Ubiquitin promoter expresses GFP and humanized spCas9 (from PX330) via a T2A motif. For cell transfection or use in lentiviral packaging. Use Pac1 and/or BstB1 sites to insert sgRNAs from PXL. FUGW
    pRubiC-T2A-Cas9Cas9Ubiquitin Luikart A Retroviral CRISPR-Cas9 System for Cellular Autism-Associated Phenotype Discovery in Developing Neurons. Sci Rep. 2016 May 10;6:25611. doi: 10.1038/srep25611. Ubiquitin promoter expresses mCherry and humanized spCas9 (PX330) via a T2A motif. For cell transfection or use in retroviral (MMuLV) packaging. Use Pac1 and/or BstB1 sites to insert sgRNAs from PXL. pRubi
    pRubiG-T2A-Cas9Cas9Ubiquitin Luikart A Retroviral CRISPR-Cas9 System for Cellular Autism-Associated Phenotype Discovery in Developing Neurons. Sci Rep. 2016 May 10;6:25611. doi: 10.1038/srep25611. Ubiquitin promoter expresses GFP and humanized spCas9 (PX330) via a T2A motif. For cell transfection or use in retroviral (MMuLV) packaging. Use Pac1 and/or BstB1 sites to insert sgRNAs from PXL. pRubi
    PXLCas9U6 Luikart A Retroviral CRISPR-Cas9 System for Cellular Autism-Associated Phenotype Discovery in Developing Neurons. Sci Rep. 2016 May 10;6:25611. doi: 10.1038/srep25611. PX330 derived. Bbs1 & annealed oligos to insert guide strand. BstB1+Pac1 of PXL and FUX-/pRubiX- T2A-Cas9 for choice of sgRNA, viral backbone, and fluorescent protein. Pac1 to introduce 2nd sgRNA. PX330
    pCas9-VRER_2A_GFPCas9-VRER (Synthetic)CAG Tessier-Lavigne Efficient introduction of specific homozygous and heterozygous mutations using CRISPR/Cas9. Nature. 2016 May 5;533(7601):125-9. doi: 10.1038/nature17664. Epub 2016 Apr 27. Cas9 VRER variant that detects NGCG PAM, combined with 2A_GFP for expression control pCAG
    pXPR_BRD111Cas9Blasticidin Hahn Integrated genetic and pharmacologic interrogation of rare cancers. NATURE COMMUNICATIONS 7:11987 Expresses Cas9 with the pLX311 backbone pLX311
    CAG-Cas9-T2A-EGFP-ires-puroWT SpCas9-T2A-EGFP-ires-puromycin resistance (Other)CAGPuromycin Otonkoski An Activating STAT3 Mutation Causes Neonatal Diabetes through Premature Induction of Pancreatic Differentiation Cell Reports 2017, 19: 281–294 Expresses WT SpCas9, EGFP and Puromycin resistance from a CAG promoter. PyCAG
    pLentiCas9-T2A-BFPT2A-BFPIn frame with Cas9Blasticidin Guigo Scalable design of paired CRISPR guide RNAs for genomic deletion (unpublished) Cas9 protein with BFP Lenti-Cas9-blast
    pLentiCas9-T2A-GFPT2A-GFPIn frame with Cas9Blasticidin Guigo Scalable design of paired CRISPR guide RNAs for genomic deletion (unpublished) Cas9 protein with GFP Lenti-Cas9-blast
    pLentiCRISPR-EPuromycin Abbosh LentiCRISPR version E (unpublished) Introduce sgRNA into a lentiviral vector (LentiCRISPR V2) which contains eSpCas9 and puromycin cassette pLentiCRISPRV2
  • Tag / Fusion Protein
    • Cas9-P2A-Puro
  • pCAG-SpCas9-GFP-U6-gRNASpCas9CAG Zou pCAG-SpCas9-U6-gRNA (unpublished) All-in-one CRISPR/Cas9 vector with CAG promoter for expression in human ESC/iPSC pSpCas9(BB)-2A-GFP (PX458)
    pCAG-eCas9-GFP-U6-gRNAeSpCas9(1.1)CAG Zou pCAG-SpCas9-U6-gRNA (unpublished) All-in-one CRISPR/Cas9 vector with high-fidelity eSpCas9 expression in human pluripotent stem cells eSpCas9(1.1)
    eSpCas9(1.1)_No_FLAGCBh Doyon A Scalable Genome-Editing-Based Approach for Mapping Multiprotein Complexes in Human Cells. Cell Rep. 2015 Oct 7. pii: S2211-1247(15)01020-7. doi: 10.1016/j.celrep.2015.09.009. FLAGless construct expressing high specificity eSpCas9(1.1). Px330-like plasmid. pX330-U6-Chimeric_BB-CBh-hSpCas9
  • Tag / Fusion Protein
    • Untagged eSpCas9(1.1)
  • pDNR-SpCas9-GemSpCas9-Gem (Other)T7 Little Cas9 nuclease fused to Geminin peptide (unpublished) Used for in vitro transcription of SpCas9-Gem mRNA. Cas9 is fused to Geminin peptide and is degraded during G0/G1 phases of the cell-cycle to minimize indels caused by non-homologous end joining pDNR-Dual
    pDNR-SpCas9SpCas9 (Other)T7 Little Cas9 nuclease fused to Geminin peptide (unpublished) Used for in vitro transcription of SpCas9 mRNA. pDNR-Dual
    pDNR-SpCas9-Cdt1SpCas9-Cdt1 (Other) Little Cas9 nuclease fused to Geminin peptide (unpublished) Used for in vitro transcription of SpCas9-Cdt1 mRNA. Cas9 is fused to Cdt1 peptide and is degraded during S/G2 phases of the cell-cycle. pDNR-Dual
    peSpCas9(1.1)-2×sgRNA (empty, donor) Nakayama Practical method for targeted disruption of cilia-related genes by using CRISPR/Cas9-mediated homology-independent knock-in system. Mol Biol Cell. 2017 Feb 8. pii: mbc.E17-01-0051. doi: 10.1091/mbc.E17-01-0051. All-in-one vector for CRISPR/Cas9-mediated homology-independent knock-in system. The plasmid contains eSpCas9(1.1) and two sgRNA expression cassettes. The first gRNA cloning site is empty. eSpCas9(1.1)
    pAAV-SMVP-Cas9NCas9N (Synthetic) Church A multifunctional AAV-CRISPR-Cas9 and its host response. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993. Expresses Cas9N in mammalian cells. pZac2.1
    pAAV-SMVP-Cas9CCas9C (Synthetic) Church A multifunctional AAV-CRISPR-Cas9 and its host response. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993. Expresses Cas9C in mammalian cells. pZac2.1
    pAAV-CASI-Cas9CCas9C (Synthetic) Church A multifunctional AAV-CRISPR-Cas9 and its host response. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993. Expresses Cas9C in mammalian cells; derived from pZac2.1 with CASI promoter. pZac2.1
    pSMVP-Cas9NCas9N (Synthetic) Church A multifunctional AAV-CRISPR-Cas9 and its host response. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993. Plasmid that expresses Cas9N in mammalian cells. pMAX
    pSMVP-Cas9C-P2A-turboGFPCas9C (Synthetic) Church A multifunctional AAV-CRISPR-Cas9 and its host response. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993. Plasmid that expresses Cas9C in mammalian cells; co-expresses turboGFP. pMAX
    pSMVP-Cas9CCas9C (Synthetic) Church A multifunctional AAV-CRISPR-Cas9 and its host response. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993. Plasmid that expresses Cas9C in mammalian cells. pMAX
    pSMVP-Cas9FL-P2A-turboGFPCas9FL (Synthetic) Church A multifunctional AAV-CRISPR-Cas9 and its host response. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993. Plasmid that expresses Cas9FL in mammalian cells; co-expresses turboGFP. pMAX
    pAAV-CASI-Cas9C-P2A-turboGFPCas9C (Synthetic) Church A multifunctional AAV-CRISPR-Cas9 and its host response. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993. Expresses Cas9C in mammalian cells; co-expresses turboGFP. pZac2.1
    pRZ-CAS9-mCherryCas9 (Synthetic)CMV Hornung CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. Cas9, mCherry expression pRZ
    pCAS9-mCherry empty Hornung CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. Cas9, mCherry, and sgRNA expression pCAS9
    pCAS9-BFP empty Hornung CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338. Cas9, TagBFP, and sgRNA expression pCAS9
    Construct 7 - CMVp-Cas9-3xNLS-HSVpACas9-3xNLS (Homo sapiens) Lu Continuous genetic recording with self-targeting CRISPR-Cas in human cells. Science. 2016 Aug 18. pii: aag0511. Expresses Cas9 fused to 3xNLS driven by CMV promoter. For mammalian cell expression pGL2-Luc (Addgene #26280)
    Construct 12 - hUBCp_Cas9_3xNLS_p2a_puroRCas9-3xNLS (Homo sapiens)Puromycin Lu Continuous genetic recording with self-targeting CRISPR-Cas in human cells. Science. 2016 Aug 18. pii: aag0511. Lentiviral construct for building stable cell lines expressing Cas9 via puromycin selection Lentiviral
    Construct 33 - NFKBRp_Cas9_3xNLS_p2a-puroRCas9-3xNLSPuromycin Lu Continuous genetic recording with self-targeting CRISPR-Cas in human cells. Science. 2016 Aug 18. pii: aag0511. Lentiviral construct for building stable cell lines expressing Cas9 in the presence of TNFa via puromycin selection Lentiviral
    LentiCRISPRv2CreCre Recombinase (Other)EFS (P2A) Feldser Systematic in vivo inactivation of chromatin regulating enzymes identifies Setd2 as a potent tumor suppressor in lung adenocarcinoma. Cancer Res. 2017 Feb 15. pii: canres.2159.2016. doi: 10.1158/0008-5472.CAN-16-2159. Lentiviral vector expressing Cre recombinase alongside Cas9 and an sgRNA cloning site lentiCRISPR v2 (Addgene #52961)
    LentiCRISPRv2GFPGFP (Other)EFS (P2A) Feldser Systematic in vivo inactivation of chromatin regulating enzymes identifies Setd2 as a potent tumor suppressor in lung adenocarcinoma. Cancer Res. 2017 Feb 15. pii: canres.2159.2016. doi: 10.1158/0008-5472.CAN-16-2159. Lentiviral vector expressing GFP alongside Cas9 and an sgRNA cloning site lentiCRISPR v2
    lentiCRISPR v2-BlastBlasticidin S deaminaseEF-1αBlasticidin Babu CRISPR/Cas9-based system for loss-of-function screening (unpublished) Mammalian expression of Cas9 and sgRNA scaffold lentiCRISPR v2 (#52961)
    pCW-Cas9-BlastBlasticidin S deaminasehuman phosphoglycerate kinase 1 promoterBlasticidin Babu CRISPR/Cas9-based system for loss-of-function screening (unpublished) Mammalian expression of doxycycline-inducible Cas9 pCW-Cas9 (#50661)
    pLV-U6-gRNA-UbC-DsRed-P2A-BsrDsRed-Express2, BsrhUbCBlasticidin Gersbach CRISPR-Cas9 epigenome editing enables high-throughput screening for functional regulatory elements in the human genome. Nat Biotechnol. 2017 Apr 3. doi: 10.1038/nbt.3853. Lentiviral SpCas9-gRNA expression vector with DsRed-Express2-P2A-BlastR FUGW
    pLV-U6-gRNA-UbC-eGFP-P2A-BsreGFP, BsrhUbCBlasticidin Gersbach CRISPR-Cas9 epigenome editing enables high-throughput screening for functional regulatory elements in the human genome. Nat Biotechnol. 2017 Apr 3. doi: 10.1038/nbt.3853. Lentiviral SpCas9-gRNA expression vector with eGFP-P2A-BlastR FUGW
    pPB-US-ECasEERT2-spCas9-ERT2 (Other)U6 (gRNA), EF1a (ERT2-Cas9-ERT2)Puromycin Chen Chen lab CRISPR plasmids (unpublished) Piggyac transposon containing tamoxifen inducible Cas9 and a gRNA cassette. Piggybac Transposon
    pX601-mCherrymCherryT2A Kan Genome editing using CRISPR-Cas9 to create the HPFH genotype in HSPCs: An approach for treating sickle cell disease and beta-thalassemia. Proc Natl Acad Sci U S A. 2016 Sep 20;113(38):10661-5. doi: 10.1073/pnas.1612075113. Epub 2016 Sep 6. Staphylococcus aureus (SaCas9) conjugated with mCherry pX601
    pX601-GFPGFP (Other) Kan Genome editing using CRISPR-Cas9 to create the HPFH genotype in HSPCs: An approach for treating sickle cell disease and beta-thalassemia. Proc Natl Acad Sci U S A. 2016 Sep 20;113(38):10661-5. doi: 10.1073/pnas.1612075113. Epub 2016 Sep 6. Staphylococcus aureus (SaCas9) conjugated with GFP pX601
    iCasCas9 (Other)CMVOFP expression Tan A chemical-inducible CRISPR-Cas9 system for rapid control of genome editing. Nat Chem Biol. 2016 Sep 12. doi: 10.1038/nchembio.2179. Expression of SpCas9 with 4 ERT2 fusion protein and empty gRNA cassette. The activity of Cas9 can be switched on and off in human cells with 4-hydroxytamoxifen (4-HT) GeneArt CRISPR Nuclease vector with OFP reporter
    pK237.U6-Chimeric-CAG-loxP-stop-loxP-hspCas9 (Supernova) loxP-stop-loxP (Other) Iwasato Supernova: A Versatile Vector System for Single-Cell Labeling and Gene Function Studies in vivo. Sci Rep. 2016 Oct 24;6:35747. doi: 10.1038/srep35747. This plasmid should be used for Cre/loxP-based Supernova-CRISPR/Cas9. pX330-U6-Chimeric_BB-CBh-hSpCas9 (Addgene Plasmid #42230)
    Lenti-iCas9-neoFlag-iCas9-P2A-GFP (Other)Neomycin (select with G418) Yan An easy and efficient inducible CRISPR/Cas9 platform with improved specificity for multiple gene targeting. Nucleic Acids Res. 2016 Nov 2;44(19):e149. Epub 2016 Jul 25. Lentiviral vector encoding a doxycycline inducible EGFP reporter downstream of FLAG-tagged spCas9, separated by a P2A self-cleavage sequence pInducer-20
    Lenti-multi-CRISPRPuromycin Yan An easy and efficient inducible CRISPR/Cas9 platform with improved specificity for multiple gene targeting. Nucleic Acids Res. 2016 Nov 2;44(19):e149. Epub 2016 Jul 25. Lentiviral vector for the delivery of multiple sgRNAs targeting different genes with constitutive Cas9 expression Lenti-CRISPR
    pAAV-RSV-SpCas9pRSV (Other)pRSV Lei The CRISPR/Cas9-created MDM2 T309G enhances vitreous-induced expression of MDM2 and proliferation and survival of cells. J Biol Chem. 2016 May 31. pii: jbc.M116.729467. Express SpCas9 in mammalian cells pAAV
    pCK002_U6-Sa-sgRNA(mod)_EFS-SaCas9-2A-Puro_WPREU6-modified-sgRNA-Backbone-EFS-hSaCas9-2xNLS-2A-Puro (Synthetic)Puromycin Regev A Distinct Gene Module for Dysfunction Uncoupled from Activation in Tumor-Infiltrating T Cells. Cell. 2016 Sep 8;166(6):1500-1511.e9. doi: 10.1016/j.cell.2016.08.052. Lentiviral vector encoding modified SaCas9 system backbone bearing BsmBI site for new guide RNAs and puromycin selection marker. pLKO
    FUCas9GFPhumanised S.pyogenes Cas9 (Other)hUBC Herold (unpublished) Expresses Cas9 with green fluorescent reporter FUGW
    pLenti-Cas9-GFPCas9-GFP (Synthetic)EFS Sabatini Gene Essentiality Profiling Reveals Gene Networks and Synthetic Lethal Interactions with Oncogenic Ras. Cell. 2017 Feb 1. pii: S0092-8674(17)30061-2. doi: 10.1016/j.cell.2017.01.013. Lentiviral vector expressing Cas9-GFP pLentiCRISPR v1
    Hygro-Cas9 donorhSpCas9 (Other)Hygromycin Huangfu Genome Editing in hPSCs Reveals GATA6 Haploinsufficiency and a Genetic Interaction with GATA4 in Human Pancreatic Development. Cell Stem Cell. 2017 Feb 8. pii: S1934-5909(17)30001-2. doi: 10.1016/j.stem.2017.01.001. Donor vector for genomic targeting of a Tetracycline-inducible Cas9 cassette to the human AAVS1/PPP1R12C locus with hygromycin selection N/A
    pU6-(BbsI)sgRNA_CAG-Cas9-venus-bpACas9-venus (Synthetic)CAG Kuehn Kuehn -sgRNA plasmids (unpublished) For cloning and expression of sgRNA together with expression of a Cas9-Venus fusion protein Bluescript
    pU6-(BbsI)sgRNA_CAG-Cas9-bpA_EF1-TagRFPCas9 (Other)CAG Kuehn Kuehn -sgRNA plasmids (unpublished) For cloning and expression of sgRNA together with expression of Cas9 and TagRFP Bluescript
    pCAG-1BPNLS-Cas9-1BPNLS1BPNLS-Cas9-1BPNLS (Synthetic)CAG Belmonte In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration. Nature. 2016 Dec 1;540(7631):144-149. doi: 10.1038/nature20565. Epub 2016 Nov 16. BPNLS-Cas9-BPNLS expression plasmid pCAG
    pCAG-1BPNLS-Cas9-1BPNLS-2AGFP1BPNLS-Cas9-1BPNLS-2A-EGFP (Synthetic)CAG Belmonte In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration. Nature. 2016 Dec 1;540(7631):144-149. doi: 10.1038/nature20565. Epub 2016 Nov 16. BPNLS-Cas9-BPNLS-2AGFP expression plasmid pCAG
    pAAV-nEFCas9nEF-Cas9 (Synthetic) Belmonte In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration. Nature. 2016 Dec 1;540(7631):144-149. doi: 10.1038/nature20565. Epub 2016 Nov 16. SpCas9 expression AAV backbone plasmid under control of nEF promoter PX551
    lenti-Cas9-VQR-BlastSpCas9-VQR(D1135V,R1335Q,T1337R) (Other)Blasticidin Bauer Variant-aware saturating mutagenesis using multiple Cas9 nucleases identifies regulatory elements at trait-associated loci. Nat Genet. 2017 Feb 20. doi: 10.1038/ng.3793. lentiviral expression of SpCas9-VQR (NGA PAM restricted) lentiCas9-Blast
    TLCV2Tight TRE promoterPuromycin Karpf Inducible LentiCRISPR v2 (unpublished) LentiCRISPR v2 was modified into an all-in-one dox inducible system. The addition of doxycycline induces Cas9-2A-eGFP. The U6 promoter drives constitutive sgRNA expression. lentiCRISPR v2
  • Tag / Fusion Protein
    • Cas9-T2A-eGFP
  • pCSDest2-NmeCas9-NLS-3XHA-NLSNme Cas9 (Other)CMV Sontheimer Naturally Occurring Off-Switches for CRISPR-Cas9. Cell. 2016 Dec 15;167(7):1829-1838.e9. doi: 10.1016/j.cell.2016.11.017. Epub 2016 Dec 8. Mammalian expression of wild type Nme Cas9 pCSDest2
    pTE4254St chimeric gRNA, hStCas9human U6, CMVNeomycin (select with G418) Welker Cpf1 nucleases demonstrate robust activity to induce DNA modification by exploiting homology directed repair pathways in mammalian cells. Biol Direct. 2016 Sep 14;11:46. doi: 10.1186/s13062-016-0147-0. Expresses St chimeric gRNA and human codon-optimized StCas9 pcDNA3.3-TOPO
    pKS7107Nm crRNA, Nm tracrRNA, hNmCas9human U6, human U6, EF1a Welker Cpf1 nucleases demonstrate robust activity to induce DNA modification by exploiting homology directed repair pathways in mammalian cells. Biol Direct. 2016 Sep 14;11:46. doi: 10.1186/s13062-016-0147-0. Expresses Nm crRNA, Nm tracrRNA and human codon-optimized NmCas9 pSimpleII

    Bacteria

    Plasmid Gene/Insert Promoter PI Publication Hidden Extra Search Info
    pMJ806Cas9T7 Doudna A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012 Jun 28. pEC-K-MBP
    pCas9tracr/Cas9 Marraffini RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2508. Bacterial expression of Cas9 nuclease, tracrRNA and crRNA guide pACYC184
    pwtCas9-bacteriawild-type Cas9pLtetO-1 Qi Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022. aTc-inducible expression of wild-type Cas9 (S .pyogenes) for bacterial gene knockdown pUC19
    pET-28b-Cas9-HisCas9 (Other) Schier Efficient mutagenesis by Cas9 protein-mediated oligonucleotide insertion and large-scale assessment of single-guide RNAs. PLoS One. 2014 May 29;9(5):e98186. doi: 10.1371/journal.pone.0098186. eCollection 2014. For in vitro expression and purification of Cas9 protein pET-28b
    DS-SPcasCas9 (Other), tracrRNA precursor (Other)proC, tracrRNA promoter Church Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. Bacterial S. pyogenes Cas9 (SP) + tracrRNA expression, cloDF13/spectinomycin cloDF13-aadA
    DS-NMcasCas9 (Other), NM tracrRNA (Other)proC, NM tracrRNA promoter Church Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. Bacterial N. meningitidis Cas9 (NM) + tracrRNA expression, cloDF13/spectinomycin cloDF13-aadA
    DS-ST1casCas9 (Other), ST1 tracrRNA (Other)proC Church Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. Bacterial S. thermophilus #1 Cas9 (ST1) + tracrRNA expression, cloDF13/spectinomycin cloDF13-aadA
    DS-TDcasCas9 (Other), TD tracrRNA (Other)proC Church Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. Bacterial T. denticola Cas9 (TD) + tracrRNA expression, cloDF13/spectinomycin cloDF13-aadA
    pET28a/Cas9-CysCas9-Cys (Other)T7 Promoter Kim Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA. Genome Res. 2014 Apr 2. Expresses N-terminal His tag fused human codon-optimized Cas9 nuclease having C-terminal Cysteine pET28a
    pCRISPomyces-1sSpCas9 (Synthetic), tracrRNA (Other), crRNA cassette (Synthetic)rpsLp(XC)-BbsI, rpsLp(CF), gapdhp(EL) Zhao High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System. ACS Synth Biol. 2014 Dec 8. Streptomyces expression of codon-optimized Cas9, tracrRNA, and custom crRNA pKC1139
    pCRISPomyces-2sSpCas9 (Synthetic), gRNA cassette (Synthetic)rpsL(XC)-BbsI, gapdhp(EL) Zhao High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System. ACS Synth Biol. 2014 Dec 8. Streptomyces expression of codon-optimized Cas9 and custom gRNA pKC1139
    pCascas9 (Other)native cas9 promoter Yang Multigene editing in the Escherichia coli genome using the CRISPR-Cas9 system. Appl Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14. Constitutive expression of cas9 and inducible expression of lambda RED and sgR pKD46
    pET-(-30)dGFP-9xGGS-Cath-NLS-Cas9-6xHis(pos36)dGFP-9xGGS-Cath-NLS-Cas9-6xHis (Other)T7 Liu Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30. Expression of (pos36)dGFP-9xGGS-Cath-NLS-Cas9-6xHis in bacterial cells pET29
    pET-Cas9-6xHisCas9_6xHis (Other)T7 Liu Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30. Expression of Cas9-6xHis in bacterial cells pET29
    pTrex-b-NLS-hSpCas9NLS-hSpCas9Ribo-HX1 Tarleton CRISPR-Cas9-Mediated Single-Gene and Gene Family Disruption in Trypanosoma cruzi. MBio. 2014 Dec 30;6(1). pii: e02097-14. doi: 10.1128/mBio.02097-14. Expression of Cas9 in T. cruzi pTrex-b
    pKCcas9dOScocas9 (Other), sgRNA (), act-orf4 homology armptipA, j23119 Jiang One-step high-efficiency CRISPR/Cas9-mediated genome editing in Streptomyces. Acta Biochim Biophys Sin (Shanghai). 2015 Apr;47(4):231-43. doi: 10.1093/abbs/gmv007. Epub 2015 Mar 3. High efficiency Streptomyces genome editing by CRISPR/Cas9 system pKC1139
    pCas9-CR4Cas9 nuclease Prather The no-SCAR (Scarless Cas9 Assisted Recombineering) system for genome editing in Escherichia coli. Sci Rep. 2015 Oct 14;5:15096. doi: 10.1038/srep15096. Cas9 nuclease under control of pTet promoter with ssrA tag and constitutive tetR pdCas9-bacteria
    SP-Cas9SP-CAS9 (Other)T7 Geijsen Efficient Intracellular Delivery of Native Proteins. Cell. 2015 Apr 23;161(3):674-690. doi: 10.1016/j.cell.2015.03.028. Expresses SP-Cas9 protein in bacterial cells pET-15
    pCMKCas9 (Synthetic) Rodrigue Bacterial constitutive gRNA expression plasmid (unpublished) Bacterial constitutive S. pyogenes Cas9 expression plasmid pCMK
    pET-Cas9-NLS-6xHisCas9-NLS-6xHis (Other)T7 Liu Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30. Expression of Cas9-NLS-6xHis in bacterial cells pET29
    pET-NLS-Cas9-6xHisNLS-Cas9-6xHis (Other)T7 Liu Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30. Expression of NLS-Cas9-6xHis in bacterial cells pET29
    pLPhygCAS9Humanized Streptococcus pyogenes Cas9 from PX330 (Addgene) (Other) Matlashewski CRISPR-Cas9-Mediated Genome Editing in Leishmania donovani. MBio. 2015 Jul 21;6(4). pii: e00861-15. doi: 10.1128/mBio.00861-15. Expresses CAS9 in Leishmania pLPHyg2
    pYTK036Cas9 (Other) Dueber A Highly-characterized Yeast Toolkit for Modular, Multi-part Assembly. ACS Synth Biol. 2015 Apr 14. Encodes Cas9 as a Type 3 part to be used in the Dueber YTK system pYTK001
    BPK764mammalian codon-optimized Streptococcus pyogenes Cas9-NLS-3XFlag, and SpCas9 gRNA (Other)T7 (x2) Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Bacterial expression plasmid for SpCas9 & sgRNA (need to clone spacer into BsaI sites): T7-humanSpCas9-NLS-3xFLAG-T7-BsaIcassette-Sp-sgRNA pACYCDuet-1
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)
  • MSP1673mammalian codon-optimized Streptococcus thermophilus1 Cas9-NLS, and St1Cas9 gRNA (Other)T7 (x2) Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Bacterial expression plasmid for S. thermophilus1 Cas9 & sgRNA (need to clone in spacer into BspMI sites): T7-humanSt1Cas9-NLS-T7-BspMIcassette-St1-sgRNA pACYCDuet-1
  • Tag / Fusion Protein
    • NLS (C terminal on insert)
  • BPK2101mammalian codon-optimized Staphylococcus aureus Cas9-NLS-3xFlag, and SaCas9 gRNA (Other)T7 (x2) Joung Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592. Bacterial expression plasmid for S. aureus Cas9 & sgRNA (need to clone in spacer into BsaI sites): T7-humanSaCas9-NLS-3xFLAG-T7-BsaIcassette-Sa-sgRNA pACYCDuet-1
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)
  • pHO4d-Cas9Cas9 (Other)T7 Nonet Landscape of target:guide homology effects on Cas9-mediated cleavage. Nucleic Acids Res. 2014 Dec 16;42(22):13778-87. doi: 10.1093/nar/gku1102. Epub 2014 Nov 15. Cas9nlsHis6 Bacterial Expression construct pHO4D
    pMJ915cas9 (Other)T7 Doudna Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery. Elife. 2014 Dec 15;3:e04766. doi: 10.7554/eLife.04766. Express Streptococcus pyogenes Cas9 carrying two C-terminal SV40 NLS pML-2CT
    MSP2283mammalian codon-optimized Staphylococcus aureus Cas9-NLS-3xFlag (Other), SaCas9 sgRNA (+84)T7, T7 Joung Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404. Bacterial expression plasmid for SaCas9 & sgRNA targeted to site 1: T7-humanSaCas9-NLS-3xFLAG-T7-Sa-sgRNA(84) #1 pACYCDuet-1
    MSP2253mammalian codon-optimized KKH variant Staphylococcus aureus Cas9(E782K/N968K/R1015H)-NLS-3xFlag (Other), SaCas9 sgRNA (+84)T7, T7 Joung Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404. Bacterial expression plasmid for KKH SaCas9 & sgRNA targeted to site 1: T7-humanSaCas9(E782K/N968K/R1015H)-NLS-3xFLAG-T7-Sa-sgRNA(84) #1 pACYCDuet-1
    MSP2266mammalian codon-optimized Staphylococcus aureus Cas9-NLS-3xFlag (Other), SaCas9 sgRNA (+84)T7, T7 Joung Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404. Bacterial expression plasmid for SaCas9 & sgRNA targeted to site 2: T7-humanSaCas9-NLS-3xFLAG-T7-Sa-sgRNA(84) #2 pACYCDuet-1
    MSP2292mammalian codon-optimized KKH variant Staphylococcus aureus Cas9(E782K/N968K/R1015H)-NLS-3xFlag, and SaCas9 sgRNA(84) (Other), SaCas9 sgRNA (+84)T7, T7 Joung Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404. Bacterial expression plasmid for KKH SaCas9 & sgRNA targeted to site 2: T7-humanSaCas9(E782K/N968K/R1015H)-NLS-3xFLAG-T7-Sa-sgRNA(84) #2 pACYCDuet-1
    pCas9_sgRNA_0U6 promoter (Other), cas9 (Synthetic), tnos terminator (Other)U6 from Ustilago maydis, synthetic otef promoter (Spellig et al., 1996, Mol. Gen. Genet. 252) Kahmann Genome editing in Ustilago maydis using the CRISPR-Cas system. Fungal Genet Biol. 2015 Sep 11. pii: S1087-1845(15)30025-6. doi: 10.1016/j.fgb.2015.09.001. expresses Ustilago maydis codon-optimized Cas9, contains U. maydis U6 promoter, is self-replicating pNEBUC
    pMCSG7-Wt-NmeCas9pMCSG7-WT-NmeCas9 (Other) Sontheimer DNase H Activity of Neisseria meningitidis Cas9. Mol Cell. 2015 Oct 15;60(2):242-55. doi: 10.1016/j.molcel.2015.09.020. bacterial pMCSG7 expression vector expressing Wildtype Nme cas9 with T7 promoter, N-terminal His tag and TEV site pMCSG7
    pCas9curCas9 (Other), Plasmid curing systempBAD Chen Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing. Metab Eng. 2015 Sep;31:13-21. doi: 10.1016/j.ymben.2015.06.006. Epub 2015 Jun 30. Constitutive expression of Cas9 gene and inducible expression of the plasmid curing system for CRISPR mediated genome editing of E. coli. pSC101ts
    pREDCas9Cas9 (Other), lambda red genes, plasmid curing systempBAD Chen Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing. Metab Eng. 2015 Sep;31:13-21. doi: 10.1016/j.ymben.2015.06.006. Epub 2015 Jun 30. Constitutive expression of Cas9, inducible expression of the Red recombineering system, and inducible expression of the plasmid curing system for CRISPR mediated genome editing of E. coli. pSC101ts
    pCAS1ylCas9 (Other), sgRNA (Synthetic)unknown Yang Multiplex gene editing of the Yarrowia lipolytica genome using the CRISPR-Cas9 system. J Ind Microbiol Biotechnol. 2016 Aug;43(8):1085-93. doi: 10.1007/s10295-016-1789-8. Epub 2016 Jun 27. Constitutive expression of Cas9 and sgRNA in Yarrowia lipolytica cells pMCSCen1
    pMA7CR_2.0cas9 (Other), lambda beta (Other), dam (Other), recX (Other)pTet, pAra, pAra, pTet Nielsen CRMAGE: CRISPR Optimized MAGE Recombineering. Sci Rep. 2016 Jan 22;6:19452. doi: 10.1038/srep19452. pMA7CR_2.0 contains arabinose inducible λ/RED β-protein and dam, and aTc inducible CRISPR/Cas9 and recX pBAD24
    pCas9-CATCas9 (Other), Chloramphenicol acetyltransferaseTgTUB1, TgTUB1 Lourido A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 10.1016/j.cell.2016.08.019. Epub 2016 Sep 2. Encodes Cas9 and chloramphenicol acetyltransferase (CAT) pU6-Universal
    pU6-DecoyDecoy sgRNAU6 Lourido A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 10.1016/j.cell.2016.08.019. Epub 2016 Sep 2. Encodes Cas9 and a CRISPR sgRNA that alleviates toxicity to Cas9 in Toxoplasma gondii pU6-Universal
    pEC-K-CBP_CjeCas9CjeCas9 (Other) Doudna Single-Stranded DNA Cleavage by Divergent CRISPR-Cas9 Enzymes. Mol Cell. 2015 Nov 5;60(3):398-407. doi: 10.1016/j.molcel.2015.10.030. Expresses Campylobacter jejuni Cas9 (Cje Cas9). pEC-K
    p2CT-CdiCdiCas9 Doudna Single-Stranded DNA Cleavage by Divergent CRISPR-Cas9 Enzymes. Mol Cell. 2015 Nov 5;60(3):398-407. doi: 10.1016/j.molcel.2015.10.030. Expresses Corynebacterium diphtheriae Cas9 (CdiCas9) PET
    pLdCNgRNA and Cas9 (Other)L. donovani ribosome RNA promoter Matlashewski Optimized CRISPR-Cas9 Genome Editing for Leishmania and Its Use To Target a Multigene Family, Induce Chromosomal Translocation, and Study DNA Break Repair Mechanisms. mSphere. 2017 Jan 18;2(1). pii: e00340-16. doi: 10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb. Express gRNA and Cas9 in Leishmania with Neomycin resistance pSP72
    pLdCHgRNA and Cas9 (Other)L. donovani ribosome RNA promoter Matlashewski Optimized CRISPR-Cas9 Genome Editing for Leishmania and Its Use To Target a Multigene Family, Induce Chromosomal Translocation, and Study DNA Break Repair Mechanisms. mSphere. 2017 Jan 18;2(1). pii: e00340-16. doi: 10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb. Express gRNA and Cas9 in Leishmania with Hygromycin resistance pSP72
    pX2-Cas9Cas9 (Other)pBAD Gill pX2-Cas9 (unpublished) Arabinose inducible Cas9 in a broad host range backbone. pBTBX-2
    pSHS212Cas9 (Other)T7 Doudna Conformational control of DNA target cleavage by CRISPR-Cas9. Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 2015 Oct 28. Cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid pCT10
    pSHS293Cas9 (Other)T7 Doudna Conformational control of DNA target cleavage by CRISPR-Cas9. Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 2015 Oct 28. D435C/E945C in cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid, lobe closure FRET construct pCT10
    pSHS306Cas9 (Other)T7 Doudna Conformational control of DNA target cleavage by CRISPR-Cas9. Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 2015 Oct 28. S867C/S355C in cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid, HNH-1 FRET construct pCT10
    pSHS248Cas9 (Other)T7 Doudna Conformational control of DNA target cleavage by CRISPR-Cas9. Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 2015 Oct 28. S867C/N1054C in cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid, HNH-2 FRET construct pCT10

    Drosophila

    Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    pHsp70-Cas9Codon optimized Cas9 O'Connor-Giles Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. Genetics. 2013 May 24. The codon-optimized Cas9 nuclease under the control of the Drosophila hsp70 promoter used in Gratz, et al. (2013). Plasmid is low copy number. pHSS6
    pBS-Hsp70-Cas9codon optimized Cas9Hsp70 O'Connor-Giles FlyCRISPR (unpublished) A codon-optimized Cas9 nuclease under the control of the Drosophila hsp70 promoter. pBluescript-KS(+)
    pAc-sgRNA-Cas9Cas9 (Synthetic), dU6-sgRNA (Drosophila melanogaster)Actin-5c, Drosophila U6Puromycin Liu Mutagenesis and homologous recombination in Drosophila cell lines using CRISPR/Cas9. Biol Open. 2013 Dec 10. pii: bio.20137120v1. doi: 10.1242/bio.20137120. Expresses sgRNA and Cas9-Puro in Drosophila S2 cells pAc-STABLE1-Puro
    pRB14S. pyogenes cas9 with humanized codon bias (Other)tubulin Foerstemann Efficient chromosomal gene modification with CRISPR/cas9 and PCR-based homologous recombination donors in cultured Drosophila cells. Nucleic Acids Res. 2014 Apr 19. expresses a myc-tagged version of hCas9 in Drosophila pCASPER5
    pDCC6Cas9 (Synthetic), U6-2>sgRNA (Synthetic)hsp70Bb, U6-96Ab Duchek Efficient CRISPR/Cas9 Plasmids for Rapid and Versatile Genome Editing in Drosophila. G3 (Bethesda). 2014 Sep 17. pii: g3.114.014126. doi: 10.1534/g3.114.014126. Bi-cistronic Drosophila CRISPR/Cas9 vector, contains hsp70>Cas9 and U6>sgRNA cassette pHW
    pnos-Cas9-noscas9nanos Bullock Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila. Proc Natl Acad Sci U S A. 2014 Jul 7. pii: 201405500. Expresses Cas9 under control of nanos promoter and 3'UTR. For germ line restricted genome engineering in Drosophila melanogaster. pnos
    pAct-Cas9cas9act5C Bullock Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila. Proc Natl Acad Sci U S A. 2014 Jul 7. pii: 201405500. Expresses Cas9 under control of act5C promoter and SV40 3'UTR. For ubiquitous genome engineering in Drosophila melanogaster. pact
    p(bhsp68-Cas9)Cas9 (Other)bhsp Averof Efficient CRISPR-mediated gene targeting and transgene replacement in the beetle Tribolium castaneum. Development. 2015 Jul 9. pii: dev.125054. Tribolium basal heat shock promoter driving Cas9 pSLfa[Tc-bhsp68::nlsEGFP]fa

    C. elegans

    Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    Peft-3::cas9-SV40_NLS::tbb-2 3'UTRcodon optimized Cas9_SV40 NLS with intron (Synthetic)eef-1A.1 (eft-3) Calarco Heritable genome editing in C. elegans via a CRISPR-Cas9 system. Nat Methods. 2013 Jun 30. doi: 10.1038/nmeth.2532. pUC57
    pDD162 (Peft-3::Cas9 + Empty sgRNA)Cas9 (Synthetic), Empty sgRNA (Other)eef-1A.1 (eft-3) , U6 Goldstein Engineering the Caenorhabditis elegans genome using Cas9-triggered homologous recombination. Nat Methods. 2013 Sep 1. doi: 10.1038/nmeth.2641. Cas9 + sgRNA plasmid that can be modified to cleave any Cas9 target site in the C. elegans genome. pCFJ601
    SP6-hCas9-Ce-mRNAhCas9 with C. elegans 5' and 3' UTRs (Homo sapiens)SP6 Sternberg Transgene-Free Genome Editing in Caenorhabditis elegans Using CRISPR-Cas. Genetics. 2013 Aug 26. In vitro transcription of a mRNA encoding humanized Cas9 nuclease, with 3' UTRs suitable for germline expression in C. elegans, for doing CRISPR-Cas by RNA injection. Uses SP6 RNA polymerase. Bluescript
    pIK86Cas9 (Synthetic)T7 Katic Targeted Heritable Mutation and Gene Conversion by Cas9-CRISPR in Caenorhabditis elegans. Genetics. 2013 Aug 26. C. elegans codon-optimized Cas9 with 2xSV40 NLS sequences. It has a T7 promoter for in vitro transcription, germline-compatible 5'UTR, tbb-2 3'UTR and a polyA sequence. pUC57
    pMB62Cas9 (Synthetic)eef-1A.1 (eft-3) Boxem CRISPR/Cas9-Targeted Mutagenesis in Caenorhabditis elegans. Genetics. 2013 Aug 26. Expresses C. elegans optimized Cas9::mEGFP from the eef-1A.1 (eft-3) promoter pBluescript SK+
    pMB63Cas9 (Synthetic)eef-1A.1 (eft-3) Boxem CRISPR/Cas9-Targeted Mutagenesis in Caenorhabditis elegans. Genetics. 2013 Aug 26. Expresses C. elegans optimized Cas9 from the eef-1A.1 (eft-3) promoter pBluescript SK+
    pMB66Cas9 (Synthetic)hsp-16.48 Boxem CRISPR/Cas9-Targeted Mutagenesis in Caenorhabditis elegans. Genetics. 2013 Aug 26. Expresses C. elegans optimized Cas9::mEGFP from the hsp-16.48 promoter pBluescript SK+
    pMB67Cas9 (Synthetic)hsp-16.48 Boxem CRISPR/Cas9-Targeted Mutagenesis in Caenorhabditis elegans. Genetics. 2013 Aug 26. Expresses C. elegans optimized Cas9 from the hsp-16.48 promoter pBluescript SK+
    peft-3::cas-9::tbb-2 3'UTRcas9eef-1A.1 (eft-3) de Bono Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination. Nucleic Acids Res. 2013 Sep 5. this plasmid contains the codon optimized gene encoding Cas9 for C. elegans. pDEST
    pJW1259Deletion Ward Rapid and Precise Engineering of the Caenorhabditis elegans Genome with Lethal Mutation Co-conversion and Inactivation of NHEJ Repair. Genetics. 2014 Dec 9. pii: genetics.114.172361. Cas9 plasmid with sgRNA deleted; for use in C. elegans pJW1219
    pME-Cas9Cas9 (Danio rerio) Zon A CRISPR/Cas9 Vector System for Tissue-Specific Gene Disruption in Zebrafish. Dev Cell. 2015 Mar 4. pii: S1534-5807(15)00075-1. doi: 10.1016/j.devcel.2015.01.032. Middle Entry clone for Gateway containing a zebrafish codon-optimized Cas9 flanked by 2 NLS pME-MCS (Tol2Kit #237)
    pME-Cas9-T2A-GFPCas9-T2A-GFP (Danio rerio) Zon A CRISPR/Cas9 Vector System for Tissue-Specific Gene Disruption in Zebrafish. Dev Cell. 2015 Mar 4. pii: S1534-5807(15)00075-1. doi: 10.1016/j.devcel.2015.01.032. Middle Entry clone for Gateway containing a zebrafish codon-optimized Cas9 flanked by 2 NLS and followed by inframe GFP. Cas9 and GFP are separated by the sequence of a T2A self-cleaving peptide. pME-MCS (Tol2Kit #237)
    pME-Cas9codon optimized Cas9 (Other) Chen Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs. Genetics. 2015 Apr 8. pii: genetics.115.176917. expresses codon optimized Cas9 in zebrafish pME-MCS
    AP56-5sgRNA for APa13 (Caenorhabditis elegans)U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans fbf-2 gene pDD162
    AP56-6sgRNA for APa13 (Caenorhabditis elegans)U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans fbf-2 gene pDD162
    AP56-7sgRNA for APa13 (Caenorhabditis elegans)U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans fbf-2 gene pDD162
    AP56-8sgRNA for APa13 (Caenorhabditis elegans)U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans fbf-2 gene pDD162
    AP54-3sgRNA for APs12 (Caenorhabditis elegans)U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans fbf-2 gene pDD162
    AP54-5sgRNA for APs12 (Caenorhabditis elegans)U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans fbf-2 gene pDD162
    APq5271sgRNA for APa4-2U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting K08F4.2 in the middle of the ORF pDD162
    APq5210sgRNA for APa4-2U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting K08F4.2 in the middle of the ORF pDD162
    AP303-3sgRNA for APs1U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 5'end of C.elegans K08F4.2 pDD162
    AP303-4sgRNA for APs1U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 5'end of C.elegans K08F4.2 pDD162
    AP332-2sgRNA for APs4U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 5'end of C.elegans K08F4.2 pDD162
    AP332-3sgRNA for APs4U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 5'end of C.elegans K08F4.2 pDD162
    AP332-4sgRNA for APs4U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 5'end of C.elegans K08F4.2 pDD162
    AP324-2sgRNA for APs6U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans K08F4.2 pDD162
    AP324-3sgRNA for APs6U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans K08F4.2 pDD162
    AP324-4sgRNA for APs6U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans K08F4.2 pDD162
    AP334-2sgRNA for APs5U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans K08F4.2 pDD162
    AP334-5sgRNA for APs5U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans K08F4.2 pDD162
    AP334-6sgRNA for APs5U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans K08F4.2 pDD162
    APCSD 53sgRNA for CSD 53U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans swan-2 pDD162
    APCSD 54sgRNA for CSD 54U6 Seydoux Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. 2014 Dec;198(4):1347-56. doi: 10.1534/genetics.114.170423. Epub 2014 Sep 23. co-expression of Cas9 and a sgRNA targeting 3'end of C.elegans swan-2 pDD162
    AP568-2sgRNA for dpy‐10 Seydoux High Efficiency, Homology-Directed Genome Editing in Caenorhabditis elegans Using CRISPR-Cas9 Ribonucleoprotein Complexes. Genetics. 2015 Sep;201(1):47-54. doi: 10.1534/genetics.115.179382. Epub 2015 Jul 17. co-expression of Cas9 and crRNA 589 target sequence pDD162
    AP568-3sgRNA for dpy‐10 Seydoux High Efficiency, Homology-Directed Genome Editing in Caenorhabditis elegans Using CRISPR-Cas9 Ribonucleoprotein Complexes. Genetics. 2015 Sep;201(1):47-54. doi: 10.1534/genetics.115.179382. Epub 2015 Jul 17. co-expression of Cas9 and crRNA 589 target sequence pDD162
    pRB1080VQR Cas9 variant (Synthetic)eef-1A.1 (eft-3) Fire Cas9 Variants Expand the Target Repertoire in Caenorhabditis elegans. Genetics. 2015 Dec 17. pii: genetics.115.185041. Expresses VQR Cas9 variant with eef-1A.1 (eft-3) promoter pCFJ601
    pRB1081dpy-10 gRNA (Synthetic)U6 Fire Cas9 Variants Expand the Target Repertoire in Caenorhabditis elegans. Genetics. 2015 Dec 17. pii: genetics.115.185041. Expresses gRNA for dpy-10 conversion using NGAG PAM and VQR Cas9 variant pRB1017
    pRB1083VRER Cas9 variant (Synthetic)eft-3 (eef-1A.1) Fire Cas9 Variants Expand the Target Repertoire in Caenorhabditis elegans. Genetics. 2015 Dec 17. pii: genetics.115.185041. Expresses VRER Cas9 variant with eef-1A.1 (eft-3) promoter pCFJ601
    pRB1084dpy-10 gRNA (Synthetic)U6 Fire Cas9 Variants Expand the Target Repertoire in Caenorhabditis elegans. Genetics. 2015 Dec 17. pii: genetics.115.185041. Expresses gRNA for dpy-10 conversion using NGCG PAM and VRER Cas9 mutant pRB1017
    pUAS:Cas9T2AGFP;U6:sgRNA1;U6:sgRNA25x UAS (Saccharomyces cerevisiae), Cas9 (Other), T2A, GFP, U6 promoter (Danio rerio) Del Bene 2C-Cas9: a versatile tool for clonal analysis of gene function. Genome Res. 2016 Mar 8. Tissue-specific knock-out in zebrafish, Cas9 and GFP driven by a UAS promoter pminiTol2
    pUAS:Cas9T2ACre;U6:sgRNA1;U6:sgRNA25xUAS, Cas9, Cre, U6 promoter Del Bene 2C-Cas9: a versatile tool for clonal analysis of gene function. Genome Res. 2016 Mar 8. Tissue-specific knock-out in zebrafish, Cas9 and Cre driven by a UAS promoter pminiTol2

    Plant

    Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    pK7WGF2::hCas9hCas9 (Synthetic)35SKanamycin Kamoun Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease. Nat Biotechnol. 2013 Aug 8;31(8):691-3. doi: 10.1038/nbt.2655. Expesses the human codon usage Cas9 nuclease (Mali et al. Science 339, 823-826, 2013) with an N-terminal GFP tag from the 35S promoter in the plant tissue pK7WGF2
    pICH41308::hCas9hCas9 (Synthetic) Kamoun Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system. Plant Methods. 2013 Oct 11;9(1):39. Level 0 hCas9 module pICH41308
    pICH47742::2x35S-5’UTR-hCas9(STOP)-NOST35Sp::hCas9 (Synthetic) Kamoun Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system. Plant Methods. 2013 Oct 11;9(1):39. Level 1 hCas9 module pICH47742
    pAGM4723::AtU6p::sgRNA2-2x35S-5′UTR::Cas9::NOST-AtU6p::sgRNA1sgRNA_PDS2-Cas9-sgRNA_PDS1 (Synthetic) Kamoun Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system. Plant Methods. 2013 Oct 11;9(1):39. Level 2 construct pAGM4723
    pBUN4113×FLAG-NLS-zCas9-NLS (Synthetic), gRNA scaffold (Synthetic)Ubi1p, OsU3pBar Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (OsU3 promoter), Bar resistance pGreen-like binary vector
    pHSN4013×FLAG-NLS-zCas9-NLS (Synthetic), gRNA scaffold (Synthetic)2×35Sp, AtU6-26pHygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Hyg resistance pGreen-like binary vector
    pRGE31Rice snoRNA U3 and dual 35S promoter Yang RNA-Guided Genome Editing in Plants Using a CRISPR-Cas System. Mol Plant. 2013 Nov;6(6):1975-83. doi: 10.1093/mp/sst119. Epub 2013 Aug 17. Expressing sgRNA and Cas9 in plants. The sgRNA was controlled by rice snoRNA U3 promoter pUGW11
    pRGEB31Rice snoRNA U3 and dual 35S promoterHygromycin Yang RNA-Guided Genome Editing in Plants Using a CRISPR-Cas System. Mol Plant. 2013 Nov;6(6):1975-83. doi: 10.1093/mp/sst119. Epub 2013 Aug 17. Binary vector derived from pRGE31. Deliver sgRNA and Cas9 into plant. by agrobacterium mediated transformation. pCAMBIA1300
    HBT-pcoCas9pcoCas9 (Synthetic)hybrid constitutive promoter 35SPPDK Sheen Multiplex and homologous recombination-mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9. Nat Biotechnol. 2013 Aug;31(8):688-91. doi: 10.1038/nbt.2654. transient expression of pcoCas9 gene in plant cells HBT-FLAG
    pFGC-pcoCas935SPPDK:pcoCas9:NOS cassette (Synthetic)constitutive 35SPPDK promoterBasta Sheen Sheen lab CRISPR plasmids (unpublished) A binary plasmid containing 35SPPDK:pcoCas9:NOS cassette for plant expression of Cas9 and multiple cloning sites for insertion of guide RNA pFGC-RCS
    pJIT163-2NLSCas9Cas9 (Other)2x35S Gao Targeted genome modification of crop plants using a CRISPR-Cas system. Nat Biotechnol. 2013 Aug;31(8):686-8. doi: 10.1038/nbt.2650. Expression of rice codon-optimized Cas9 in plant cells pJIT163
    p201N Cas9Cas9 (Synthetic), nptII (Other)2x35S, StUbi-3PG418, kanamycin Parrott Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 2015;15:16 Cas9 driven by double 35S, nptII for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle pPZP
    p201H Cas9Cas9 (Synthetic), hph (Other)2x35S, StUbi-3PHygromycin Parrott Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 2015;15:16 Cas9 driven by double 35S, hygromycin resistance for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle pPZP
    p201B Cas9Cas9 (Synthetic), BAR (Other)2x35S, 2x35SBasta Parrott Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 2015;15:16 Cas9 driven by double 35S, BAR for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle pPZP
    p201G Cas9Cas9 (Synthetic), sGFP (Synthetic)2x35S, 2x35SGFP Parrott Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 2015;15:16 Cas9 driven by double 35S, GFP for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle pPZP
    Cas9 MDC123Glycine Max codon optimized Cas9 (Other)2X35SBasta Stupar CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a new web-tool and a modified Cas9 enzyme. GM Crops Food. 2015;6(4):243-52. doi: 10.1080/21645698.2015.1106063. 2x35S promoting Glycing Max codon optimized Cas9 with bar plant selection pMDC123
    Cas9 MDC32Glycine Max codon optimized Cas9 (Other)2X35SHygromycin Stupar CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a new web-tool and a modified Cas9 enzyme. GM Crops Food. 2015;6(4):243-52. doi: 10.1080/21645698.2015.1106063. 2x35S promoting Glycing Max codon optimized Cas9 with hygromycin plant selection pMDC32
    G10 Cas9 MDC123Glycine Max codon optimized Cas9 (Other)G10Basta Stupar CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a new web-tool and a modified Cas9 enzyme. GM Crops Food. 2015;6(4):243-52. doi: 10.1080/21645698.2015.1106063. G10 promoting Glycing Max codon optimized Cas9 (N and C terminus NLS) with bar plant selection pMDC123
    pBUE411gRNA scaffold (Synthetic), zCas9 (Other)OsU3, UbiBasta Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (OsU3 promoter), Bar resistance pCambia
    pHSE401gRNA scaffold (Synthetic), zCas9 (Other)AtU6-26p, 35SHygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Hyg resistance pCambia
    pKSE401gRNA scaffold (Synthetic), zCas9 (Other)AtU6-26p, 35SNeomycin (select with G418) Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Kan resistance pCambia
    pHUE411gRNA scaffold (Synthetic), zCas9 (Other)OsU3, UbiHygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (OsU3 promoter), Hyg resistance pCambia
    pBUN421gRNA scaffold (Synthetic), zCas9 (Other)TaU3, UbiBasta Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (TaU3 promoter), Bar resistance pGreen-like
    pRGEB32Rice snoRNA U3 promoter for gRNA/PTG expression and UBI promoter for Cas9 expressionHygromycin Yang Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice ubiquitin promoter for Agrobacterium-mediated transformation. pRGEB31
    pRGE32Rice snoRNA U3 promoter for gRNA/PTG expression and UBI promoter for Cas9 expression Yang Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice ubiquitin promoter for transient expression in protoplast pRGE31
    pYLCRISPR/Cas9Pubi-HCas9 (Other)maize ubiquitin promoter (Pubi)Hygromycin Liu A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. expression of Cas9 in plants, maize ubiquitin promoter (Pubi), hygro selection pCAMBIA1300
    pYLCRISPR/Cas9Pubi-BCas9 (Other)maize ubiquitin promoter (Pubi)Basta Liu A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. expression of Cas9 in plants, maize ubiquitin promoter (Pubi), basta selection pCAMBIA1300
    pYLCRISPR/Cas9P35S-BCas9 (Other)cauliflower mosaic virus 35S promoter (P35S)Basta Liu A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. expression of Cas9 in plants, cauliflower mosaic virus 35S promoter (P35S), basta selection pCAMBIA1300
    pEGB2alpha2 35s:hCas9:tNos (GB0639)hCas9 (Other)35S Orzaez GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. Plant Physiol. 2013 Jul;162(3):1618-31. Transcriptional unit for (human codon optimized) Cas9 plant expression driven by the 35S promoter pDGB2alpha2
    pICSL11056Promoter/5UTR: ZmUbi + CDS:SpCas9 + 3UTR/terminator:35s (Other) Patron Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease. Genome Biol. 2015 Nov 30;16(1):258. doi: 10.1186/s13059-015-0826-7. Level 1 Golden Gate Cassette: Cas9 expression cassette for monocotyledonous plants pICH47742 (AddGene #47742)
    pICSL11060Promoter/5UTR: CsVMV+ CDS:SpCas9 + 3UTR/terminator:35s (Other) Patron Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease. Genome Biol. 2015 Nov 30;16(1):258. doi: 10.1186/s13059-015-0826-7. Level 1 Golden Gate Cassette: Cas9 expression cassette for plants (exemplified in Brassica oleracea) pICH47742 (AddGene #47742)
    pYPQ150pcoCas9 (plant codon-optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with pcoCas9 (Plant codon optimized) pNJB91
    pYPQ154AteCas9 (Arabidopsis codon optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with AteCas9 (Arabidopsis codon optimized) pYPQ185-linker2
    pYPQ158hSpCas9 (Human codon optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with hSpCas9 (Human codon optimized) pNJB185-linker6
    pYPQ167Cas9p (Plant codon-optimized; high GC content at 5 prime region) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with Cas9p pYPQ185-linker2
    pTC217Nuclease (Cas9/sgRNA) + Donor + GVR (Other)kanamycin Voytas High-frequency, precise modification of the tomato genome. Genome Biol. 2015 Nov 6;16(1):232. doi: 10.1186/s13059-015-0796-9. Cas9/sgRNA targeting ANT1 locus, donor molecule with 5' homology arm-Pnos:NptII-35S:ANT13' homology arm as a Gemini Viral Replicon based on Bean Yellow Dwarf Virus; sgRNA= 1b pLSLR
    pTC223Nuclease (Cas9/sgRNA) + Donor + GVR (Other)kanamycin Voytas High-frequency, precise modification of the tomato genome. Genome Biol. 2015 Nov 6;16(1):232. doi: 10.1186/s13059-015-0796-9. Cas9/sgRNA targeting ANT1 locus, donor molecule with 5' homology arm-Pnos:NptII-35S:ANT13' homology arm as a Gemini Viral Replicon based on Bean Yellow Dwarf Virus; sgRNA= 7 pLSLR
    pHEE401sgRNA scaffold (Synthetic), zCas9 (Other)U6-26p Arabidopsis U6 gene promoter, EC1.2 enhancer fused to EC1.1 promoterHygromycin Chen Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. Egg cell-specific promoter-controlled expression of 3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target sequence (U6-26 promoter), Hyg resistance pCambia
    pHEE401EgRNA scaffold (Synthetic), zCas9 (Other)U6-26p Arabidopsis U6 gene promoter, EC1.2 enhancer fused to EC1.1 promoterHygromycin Chen Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. Egg cell-specific promoter-controlled expression of 3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target sequence (U6-26 promoter), Hyg resistance pCambia
    pHEE2E-TRIsgRNA targeting TRY and CPC genes (Synthetic), sgRNA targeting ETC2 gene (Synthetic), zCas9 (Other)U6-26p Arabidopsis U6 gene promoter, U6-26p Arabidopsis U6 gene promoter, EC1.2 enhancer fused to EC1.1 promoterHygromycin Chen Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. Egg cell-specific promoter-controlled expression 3×FLAG-NLS-zCas9-NLS and two sgRNAs targeting three genes: ETC2, TRY, and CPC pCambia
    pMpGE010Hygromycin Kohchi pMpGE010 (unpublished) For CRISPR/Cas9 genome editing in Marchantia polymorpha pMpGWB103
    pMpGE011Chlorsulfuron Kohchi pMpGE010 (unpublished) For CRISPR/Cas9 genome editing in Marchantia polymorpha pMpGWB303
    pEGB2Ω2_SF-35s:hCas9:tNos (GB1103)hCas9 (Other)35S Orzaez A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. eCollection 2016. Transcriptional unit for (human codon optimized) Cas9 plant expression driven by the 35S promoter. Specially conceived to be linked with the TU of the kanamycin resistance gene GB1181. pDGB2omega2
    pBAtCU6Basta Kim A simple, flexible and high-throughput cloning system for plant genome editing via CRISPR-Cas system. J Integr Plant Biol. 2016 Mar 4. doi: 10.1111/jipb.12474. Plant expression of Cas9 and gRNA empty backbone; Basta resistance pB2GW7
  • Tag / Fusion Protein
    • NLS-HA (C terminal on insert)
  • pHAtCU6Hygromycin Kim A simple, flexible and high-throughput cloning system for plant genome editing via CRISPR-Cas system. J Integr Plant Biol. 2016 Mar 4. doi: 10.1111/jipb.12474. Plant expression of Cas9 and gRNA empty backbone; Hygro Resistance pH2GW7
    pHDE-35S-Cas9-mCherryHygromycin ; mCherry fluorescence Zhao An effective strategy for reliably isolating heritable and Cas9-free Arabidopsis mutants generated by CRISPR/Cas9-mediated genome editing. Plant Physiol. 2016 May 15. pii: pp.00663.2016. For CRISPR/Cas9 mediated gene editing in Arabidopsis; provides a visual screen for Cas9-free plants pHDE
    pHDE-35S-Cas9-mCherry-UBQHygromycin ; mCherry fluorescence Zhao An effective strategy for reliably isolating heritable and Cas9-free Arabidopsis mutants generated by CRISPR/Cas9-mediated genome editing. Plant Physiol. 2016 May 15. pii: pp.00663.2016. For CRISPR/Cas9 mediated gene editing in Arabidopsis; provides a visual screen for Cas9-free plants. Enable production of two gRNAs pHDE
    pKIR1.1human-codon-optimized SpCas9 (Other)AtRPS5AHygromycin ; TagRFP in seeds Higashiyama pKAMA-ITACHI vectors for highly efficient CRISPR/Cas9-mediated gene knockout in Arabidopsis thaliana. Plant Cell Physiol. 2016 Nov 17. pii: pcw191. CRISPR/Cas9 in Arabidopsis with seed a fluorescent reporter pFAST-R01
    pKI1.1Rhuman-codon-optimized SpCas9 (Other)AtRPS5AHygromycin ; TagRFP in seeds Higashiyama pKAMA-ITACHI vectors for highly efficient CRISPR/Cas9-mediated gene knockout in Arabidopsis thaliana. Plant Cell Physiol. 2016 Nov 17. pii: pcw191. CRISPR/Cas9 in Arabidopsis with seed a fluorescent reporter. pFAST-R01
    pAGM4723:TpCC_UreaseCas9 (Other), Urease-targeting gRNA 1 (Other), Urease-targeting gRNA 2 (Other)FCP, U6, U6Nourseothricin (Nat) Mock Editing of the urease gene by CRISPR-Cas in the diatom Thalassiosira pseudonana Plant Methods 2016, 12:49 Golden Gate Level 2 construct encoding Cas9-YFP and 2 urease-targeting gRNAs pAGM4723
    pICH47742:FCP:Cas9YFPCas9 (Other)FCP Mock Editing of the urease gene by CRISPR-Cas in the diatom Thalassiosira pseudonana Plant Methods 2016, 12:49 Golden Gate Level 1 cassette encoding a Cas9-YFP fusion under the FCP promoter pICH47742
    pKIR1.0human-codon-optimized SpCas9 (Other)AtRPS5ATagRFP in seeds Higashiyama pKAMA-ITACHI vectors for highly efficient CRISPR/Cas9-mediated gene knockout in Arabidopsis thaliana. Plant Cell Physiol. 2016 Nov 17. pii: pcw191. CRISPR/Cas9 in Arabidopsis with seed a fluorescent reporter. pFAST-R01

    Yeast

    Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    p414-TEF1p-Cas9-CYC1tHuman Optimized S. pyogenes Cas9 (Homo sapiens)TEF1 promoterTRP1 Church Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Nucleic Acids Res p414
    p415-GalL-Cas9-CYC1tHuman Optimized S. pyogenes Cas9 (Homo sapiens)GalLLEU2 Church Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Nucleic Acids Res p415
    pACT2-CAS9cCas9c (Synthetic)yeast ADH1 promoterLEU2 Zhao Self-processing of ribozyme-flanked RNAs into guide RNAs in vitro and in vivo for CRISPR-mediated genome editing. J Integr Plant Biol. 2013 Dec 30. doi: 10.1111/jipb.12152. Express Eukaryotic-codon-optimized Cas9c gene in yeast under ADH1 promoter for genome editing. SV40 NLS is fused to the C terminal. GAL4 region from pACT2 is removed. pACT2
    pMZ222Cas9 (Other)adh1LEU2 Zaratiegui Implementation of the CRISPR-Cas9 system in fission yeast. Nat Commun. 2014 Oct 29;5:5344. doi: 10.1038/ncomms6344. Constitutive expression of humanized Cas9 pART1
    pMZ374Cas9 (Other), rrk1:sgRNA (Other)adh1, rrk1ura4 Zaratiegui Implementation of the CRISPR-Cas9 system in fission yeast. Nat Commun. 2014 Oct 29;5:5344. doi: 10.1038/ncomms6344. Combination adh1:cas9/rrk1:sgRNA for CRISPR genome editing in fission yeast: Empty sgRNA target (CspCI placeholder) (see comments). pMZ283
    pCTiCas9 (Other), tracrRNA (Other)TEF1p, RPR1pLEU2 Zhao Homology-Integrated CRISPR-Cas (HI-CRISPR) System for One-Step Multigene Disruption in Saccharomyces cerevisiae. ACS Synth Biol. 2014 Sep 19. Plasmid encoding iCas9 and tracrRNA on pRS415 backbone pRS415
    pCRCTiCas9 (Other), tracrRNA (Other)TEF1p, RPR1pURA3 Zhao Homology-Integrated CRISPR-Cas (HI-CRISPR) System for One-Step Multigene Disruption in Saccharomyces cerevisiae. ACS Synth Biol. 2014 Sep 19. Plasmid encoding iCas9, tracrRNA and crRNAs pRS426
    Cas9-NATHuman-optimized S. pyogenes Cas9 with Clonnat marker gene expression cassetteNourseothricin(clonNat) Jin Construction of a quadruple auxotrophic mutant of an industrial polyploid saccharomyces cerevisiae strain by using RNA-guided Cas9 nuclease. Appl Environ Microbiol. 2014 Dec;80(24):7694-701. doi: 10.1128/AEM.02310-14. Epub 2014 Oct 3. Cas9 expression cassette carried by a yeast single-copy episome plasmid pRS414-TEF1p-Cas9-CYC1t
    pML104Cas9 (Saccharomyces cerevisiae), single guide RNA expression cassette (Saccharomyces cerevisiae)pTDH3, pSNR52URA3 Wyrick New vectors for simple and streamlined CRISPR-Cas9 genome editing in Saccharomyces cerevisiae. Yeast. 2015 Dec;32(12):711-20. doi: 10.1002/yea.3098. Epub 2015 Sep 21. Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains URA3 marker for yeast transformation pRSII426
    pML107Cas9 (Saccharomyces cerevisiae), single guide RNA expression cassette (Saccharomyces cerevisiae)pTDH3, pSNR52LEU2 Wyrick New vectors for simple and streamlined CRISPR-Cas9 genome editing in Saccharomyces cerevisiae. Yeast. 2015 Dec;32(12):711-20. doi: 10.1002/yea.3098. Epub 2015 Sep 21. Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains LEU2 marker for yeast transformation pRSII425
    pJH001Cas9 (Saccharomyces cerevisiae), EmptyLEU2 Wyrick New vectors for simple and streamlined CRISPR-Cas9 genome editing in Saccharomyces cerevisiae. Yeast. 2015 Dec;32(12):711-20. doi: 10.1002/yea.3098. Epub 2015 Sep 21. High copy Cas9 expression vector with LEU2 marker for yeast transformation pRSII425
    p415-PtrpC-Cas9-TtrpC-CYC1thumanized Cas9 (Homo sapiens)LEU2 Hong Efficient gene editing in Neurospora crassa with CRISPR technology Fungal Biology and Biotechnology 2015, 2:4 Express humanized Cas9 under the control of trpC promoter and terminator from A. nidulans. pRS415
    pCRISPRylCodon optimized Cas9 from S. pyogenes (Synthetic), sgRNA expression cassette (Synthetic)UAS1B8-TEF(136), SCR1'-tRNALEU2 Wheeldon Synthetic RNA polymerase III promoters facilitate high efficiency CRISPR-Cas9 mediated genome editing in Yarrowia lipolytica. ACS Synth Biol. 2015 Dec 29. CRISPR/Cas9 vector for Yarrowia lipolytica, with AvrII site for sgRNA insertion pUC19
    pMZ376Cas9 (Other), rrk1:sgRNA (Other)adh1, rrk1Neomycin (select with G418) Zaratiegui sgRNA/Cas9 (unpublished) Combined sgRNA/Cas9 vector with KanMX marker pMZ374
    pMZ377Cas9 (Other), rrk1:sgRNA (Other)adh1, rrk1LEU2 Zaratiegui sgRNA/Cas9 (unpublished) Combined sgRNA/Cas9 vector with LEU2 marker pMZ374
    pMZ379Cas9 (Other), rrk1:sgRNA (Other)adh1, rrk1nourseothricin Zaratiegui sgRNA/Cas9 (unpublished) Combined sgRNA/Cas9 vector with NatMX marker pMZ374
    pCfB2312 (TEF1p-Cas9-CYC1t_kanMX)Cas9 (Other)Gentamicin Borodina EasyClone-MarkerFree: A vector toolkit for marker-less integration of genes into Saccharomyces cerevisiae via CRISPR-Cas9. Biotechnol J. 2016 Aug;11(8):1110-7. doi: 10.1002/biot.201600147. Epub 2016 Jun 23. Expresses Cas9 protein, under the control of the TEF1 promoter, and CYC terminator. KanMX resistance pRS414
    pRS416gT-GalL-Cas9Cas9 (Synthetic), guide RNA (gRNA) (Synthetic), Tetracycline Repressor (Other)GalL, RPR1 promoter with TetO site, GPM1 promoterURA3 Davis Distinct patterns of Cas9 mismatch tolerance in vitro and in vivo. Nucleic Acids Res. 2016 May 19. pii: gkw417. pRS416 (URA) + RPR1(TetO) promoter with gRNA locus + Tetracycline Repressor + GalL promoter driven Cas9 for yeast expression pRS416
    pRCC-KCas9 (Other), empty gRNA cassette (Synthetic)ROX3, SNP52pNeomycin (select with G418) Boles Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. Expression of Cas9 and gRNA cassette in S. cerevisiae; Kan Resistance pRS42K
  • Tag / Fusion Protein
    • NTS (C terminal on backbone)
  • pRCC-NCas9 (Other), empty gRNA cassette (Synthetic)ROX3, SNP52pNourseothricin Boles Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. Expression of Cas9 and gRNA cassette in S. cerevisiae; Nourseothricin Resistance pRS42N
  • Tag / Fusion Protein
    • NTS (C terminal on backbone)
  • pDuRCC-KCas9 (Other), empty gRNA cassette (Synthetic), empty gRNA cassette (Synthetic)ROX3, SNP52p, SNP52pNeomycin (select with G418) Boles Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. Expression of Cas9 and two gRNA cassettes in S. cerevisiae; Kan Resistance pRS42K
  • Tag / Fusion Protein
    • NTS (C terminal on backbone)
  • pDuRCC-NCas9 (Other), empty gRNA cassette (Synthetic), empty gRNA cassette (Synthetic)ROX3, SNP52p, SNP52pNourseothricin Boles Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. Expression of Cas9 and two gRNA cassettes in S. cerevisiae; Nourseothricin Resistance pRS42N
  • Tag / Fusion Protein
    • NTS (C terminal on backbone)
  • pML104-HygMx4HphMX4 (Other)Hygromycin Wittkopp Patricia Wittkopp yeast CRISPR plasmids (unpublished) Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains HphMx4 marker for yeast transformation. pRSII426
    pML104-KanMx4KanMX4 (Other)Neomycin (select with G418) Wittkopp Patricia Wittkopp yeast CRISPR plasmids (unpublished) Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains KanMx4 marker for yeast transformation. pRSII426
    pML104-NatMx3NatMx3 (Other)nourseothricin (nat) Wittkopp Patricia Wittkopp yeast CRISPR plasmids (unpublished) Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains NatMX3 marker for yeast transformation. pRSII426
    pCfB2312 (TEF1p-Cas9-CYC1t_kanMX)Cas9 (Other)G418 Borodina CRISPR–Cas system enables fast and simple genome editing of industrial Saccharomyces cerevisiae strains Metab Eng Commun 2015 Cas9-carrying vector with kanMX marker p414-TEF1p-Cas9-CYC1t
    pCRISPRyl_AXPCas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at AXP locus in Y. lipolytica pUC19
    pCRISPRyl_XPR2Cas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at XPR2 locus in Y. lipolytica pUC19
    pCRISPRyl_A08Cas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at A08 locus in Y. lipolytica pUC19
    pCRISPRyl_D17Cas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at D17 locus in Y. lipolytica pUC19
    pCRISPRyl_MFE1Cas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at MFE1 locus in Y. lipolytica pUC19
    pJB172gRNA targeting pil1 (Schizosaccharomyces pombe)Fluoride Berro Use of a fluoride channel as a new selection marker for fission yeast plasmids and application to fast genome editing with CRISPR/Cas9. Yeast. 2016 Oct;33(10):549-557. doi: 10.1002/yea.3178. Epub 2016 Sep 7. CRISPR/Cas9 in fission yeast using fluoride selection and targetting pil1 pJB166

    Zebrafish

    Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    MLM3613Streptococcus pyogenes Cas9CMV Joung Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2501. Expresses Cas9 nuclease (Streptococcus pyogenes) from CMV and T7 promoters unknown
    MLM3639Streptococcus pyogenes Cas9-3X FlagCMV Joung Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2501. unknown
    pST1374-NLS-flag-linker-Cas9Human codon optimized Cas9CMVBlasticidin Huang Generation of gene-modified mice via Cas9/RNA-mediated gene targeting. Cell Res. 2013 Apr 2. doi: 10.1038/cr.2013.46. pST1374 (Addgene #13426)
    pT3TS-nCas9nCas9 (Synthetic)T3 Chen Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system. Proc Natl Acad Sci U S A. 2013 Aug 5. pT3T7
    pCS2-Cas9Cas9 (Other)SP6 Schier Efficient mutagenesis by Cas9 protein-mediated oligonucleotide insertion and large-scale assessment of single-guide RNAs. PLoS One. 2014 May 29;9(5):e98186. doi: 10.1371/journal.pone.0098186. eCollection 2014. For in vivo expression of Cas9 or generation of Cas9 mRNA pCS2
    pCS2-nCas9nnls-zcas9-nls (Synthetic)CMV Chen Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system. Proc Natl Acad Sci U S A. 2013 Aug 5. expression of an optimized Cas9 for genome-editing in zebrafish pCS2+
    pCS2+hSpCas9humanized S.pyogenes Cas9SP6 Kinoshita Kinoshita lab Cas9 plasmids (unpublished) Encode hSpCas9 for in vitro transcription with Sp6 pCS2
    pCS2-nCas9n-nanos 3’UTRnanos 3' UTR (Danio rerio)SP6 Giraldez CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo. Nat Methods. 2015 Aug 31. doi: 10.1038/nmeth.3543. expression of a germ-line targeted, codon optimized Cas9 for genome-editing in zebrafish pCS2-nCas9n nanos3 cb725, nanos, nanos1, nos1
    pME-Cas9Cas9 (Danio rerio) Zon A CRISPR/Cas9 Vector System for Tissue-Specific Gene Disruption in Zebrafish. Dev Cell. 2015 Mar 4. pii: S1534-5807(15)00075-1. doi: 10.1016/j.devcel.2015.01.032. Middle Entry clone for Gateway containing a zebrafish codon-optimized Cas9 flanked by 2 NLS pME-MCS (Tol2Kit #237)
    pME-Cas9-T2A-GFPCas9-T2A-GFP (Danio rerio) Zon A CRISPR/Cas9 Vector System for Tissue-Specific Gene Disruption in Zebrafish. Dev Cell. 2015 Mar 4. pii: S1534-5807(15)00075-1. doi: 10.1016/j.devcel.2015.01.032. Middle Entry clone for Gateway containing a zebrafish codon-optimized Cas9 flanked by 2 NLS and followed by inframe GFP. Cas9 and GFP are separated by the sequence of a T2A self-cleaving peptide. pME-MCS (Tol2Kit #237)
    pME-Cas9codon optimized Cas9 (Other) Chen Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs. Genetics. 2015 Apr 8. pii: genetics.115.176917. expresses codon optimized Cas9 in zebrafish pME-MCS
    pUAS:Cas9T2AGFP;U6:sgRNA1;U6:sgRNA25x UAS (Saccharomyces cerevisiae), Cas9 (Other), T2A, GFP, U6 promoter (Danio rerio) Del Bene 2C-Cas9: a versatile tool for clonal analysis of gene function. Genome Res. 2016 Mar 8. Tissue-specific knock-out in zebrafish, Cas9 and GFP driven by a UAS promoter pminiTol2
    pUAS:Cas9T2ACre;U6:sgRNA1;U6:sgRNA25xUAS, Cas9, Cre, U6 promoter Del Bene 2C-Cas9: a versatile tool for clonal analysis of gene function. Genome Res. 2016 Mar 8. Tissue-specific knock-out in zebrafish, Cas9 and Cre driven by a UAS promoter pminiTol2
    pMJ922SpCas9 (Other)T7 Jinek Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes. Development. 2016 Apr 29. pii: dev.134809. Expression of His6-MBP-tagged Cas9-NLS-EGFP protein in E. coli pET His6 MBP N10 TEV LIC cloning vector (2C-T)
    pMJ923SpCas9 (Other)T7 Jinek Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes. Development. 2016 Apr 29. pii: dev.134809. Expression of His6-MBP-tagged Cas9-NLS-mCherry protein in E. coli pET His6 MBP N10 TEV LIC cloning vector (2C-T)

    Xenopus

    Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    pCS2-3xFLAG-NLS-SpCas9-NLS3xFLAG-NLS-SpCas9-NLSCMV Chen Efficient RNA/Cas9-mediated genome editing in Xenopus tropicalis. Development. 2014 Jan 8. Cas9 expression plasmid pCS2+