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  • CRISPR
  • Bacteria

    • CRISPR Plasmids: Bacteria

    Browse CRISPR Plasmids

    By Function
    Genome Editing
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    CRISPR Resources

    The following CRISPR plasmids have been designed for use in bacteria.

    Cut

    Fully functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a 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. Indels often lead to frameshifts, creating loss of function alleles.

    To introduce specific genomic changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead of NHEJ. In most experimental systems, HDR occurs at a much lower efficiency than NHEJ.

    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
    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(-30)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 (-30)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
    pDEST-hisMBP-AsCpf1-ECAsCpf1 (Synthetic)tac promoter Kim Targeted mutagenesis in mice by electroporation of Cpf1 ribonucleoproteins. Nat Biotechnol. 2016 Jun 6. doi: 10.1038/nbt.3596. Expressing his-MBP tagged AsCpf1 (E.coli codon optimized) pDEST-hisMBP
    pMAL-his-LbCpf1-ECLbCpf1 (Synthetic)tac promoter Kim Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol. 2016 Jun 6. doi: 10.1038/nbt.3609. Bacterial expression - MBP-his tagged LbCpf1 (E.coli codon optimized) pMAL-c5X
    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
    bbCas9pluspAAACas9 (Other)T7 Huijbers Direct Generation of Conditional Alleles Using CRISPR/Cas9 in Mouse Zygotes. Methods Mol Biol. 2017;1642:21-35. doi: 10.1007/978-1-4939-7169-5_2. The plasmid encodes the T7 promoter, Cas9 (from pX330 #42230) and a stretch of poly-A. After linearization with restriction enzyme SapI It is used to produce in vitro transcribed mRNA pUC
    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
    pJYS3_ΔcrtYfCpf1 (Other), crRNA of crtYf (Synthetic) Yang CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum. Nat Commun. 2017 May 4;8:15179. doi: 10.1038/ncomms15179. Constitutive transcription of FnCpf1 and crRNA of crtYf pXMJ19
    pJYS1PtacCpf1 (Other), recT (Other)tac, unknown Yang CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum. Nat Commun. 2017 May 4;8:15179. doi: 10.1038/ncomms15179. Constitutive trancription of FnCpf1 and recT in C.glutamitum, tac promoter pXMJ19
    pJYS1PeftuCpf1 (Other), recT (Other)etfu, unknown Yang CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum. Nat Commun. 2017 May 4;8:15179. doi: 10.1038/ncomms15179. Constitutive expression of FnCpf1 and recT in C.glutamitum, etfu promoter pXMJ19
    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
    pSHS306 - Bacterial expression plasmid for SpCas9, HNH FRET variantCas9 (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
    pET-MBP-Geo_stGeoCas9 (Other) Doudna GeoCas9 Plasmids (unpublished) Expression plasmid for Cas9 from Geobacillus stearothermophilus with an N-Term MBP pET
    pET-MBP-NLS-Geo_stGeoCas9 (Other) Doudna GeoCas9 Plasmids (unpublished) Expression plasmid for Cas9 from Geobacillus stearothermophilus with an N-Term MBP and SV40 NLS pET
    pFC330Cas9 (Synthetic), pyrG (Other)Aspergillus nidulans tef1 promoter, native promoter Mortensen A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015. AMA1 plasmid with Aspergillus optimized Cas9 and pyrG selection marker custom
    pFC331Cas9 (Synthetic), argB (Other)Aspergillus nidulans tef1 promoter, native promoter Mortensen A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015. AMA1 plasmid with Aspergillus optimized Cas9 and argB selection marker custom
    pFC333Cas9 (Synthetic), ble (bleomycin resistance marker) (Other)Aspergillus nidulans tef1 promoter, Aspergillus nidulans trpC promoter Mortensen A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015. AMA1 plasmid with Aspergillus optimized Cas9 and ble selection marker custom
    pFC332Cas9 (Synthetic), hph (hygromycin resistance marker) (Other)Aspergillus nidulans tef1 promoter, Aspergillus nidulans trpC promoter Mortensen A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015. AMA1 plasmid with Aspergillus optimized Cas9 and hph selection marker custom
    pMJ915v2+Nterm Spe1 +Cterm Age1 site Doudna Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. For expression of modified SpyCas9 in e.coli. pMJ915
    1xNLS-pMJ915v2Cas9 Doudna Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. For expression of modified SpyCas9 in e.coli. pMJ915
    2xNLS-pMJ915v2Cas9, T7 Doudna Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. For expression of modified SpyCas9 in e.coli. pMJ915
    4xNLS-pMJ915v2Cas9, T7 Doudna Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. For expression of modified SpyCas9 in e.coli. pMJ915
    pMJ915v2 + sfGFPCas9, T7 Doudna Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. For expression of modified SpyCas9 in e.coli. pMJ915
    1xNLS-pMJ915v2-sfGFPCas9, T7 Doudna Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. For expression of modified SpyCas9 in e.coli. pMJ915
    2xNLS-pMJ915v2-sfGFPCas9, T7 Doudna Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. For expression of modified SpyCas9 in e.coli. pMJ915
    4xNLS-pMJ915v2-sfGFPCas9, T7 Doudna Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. For expression of modified SpyCas9 in e.coli. pMJ915
    7xNLS-pMJ915v2-sfGFPCas9, T7 Doudna Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806. For expression of modified SpyCas9 in e.coli. pMJ915
    pET-CjCas9CjCas9 (Other)T7 Kim In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni. Nat Commun. 2017 Feb 21;8:14500. doi: 10.1038/ncomms14500. Expression of CjCas9 with His tag in E.coli pET28b
    pEM-Cas9HF1Cas9HF1 (Other)pTet Lynch Managing the SOS Response for Enhanced CRISPR-Cas-Based Recombineering in E. coli through Transient Inhibition of Host RecA Activity. ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174. Modified from pCas9-CR4 (Addgene: 62655) to use the high-fidelity version of Cas9, SpCas9-HF1 (N497A/R661A/Q695A/Q926A) from Kleinstiver et al 2016. pCas9-CR4
    pEM-Cas9HF1-recA56Cas9HF1 (Other), recA56 (Other)pTet, proD Lynch Managing the SOS Response for Enhanced CRISPR-Cas-Based Recombineering in E. coli through Transient Inhibition of Host RecA Activity. ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174. Modified from pEM-Cas9HF1 (Addgene ID: 89961) to include constitutive recA56 to block recA-mediated double-strand break repair. pEM-Cas9HF1
    6-His-MBP-TEV-FnCpf1FnCpf1 (humanized) (Other)T7 Zhang Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038. Bacterial expression plasmid for protein purification pET-28
    6His-MBP-TEV-huAsCpf1huAsCpf1 (Other) Zhang Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5. Bacterial expression plasmid for protein purification pET-28
    6His-MBP-TEV-huLbCpf1huLbCpf1 (Other) Zhang Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5. Bacterial expression plasmid for protein purification pET-28
    pMST665_BB3_L 23_gRNAempty_cas9_BbsICas9 Sauer An efficient tool for metabolic pathway construction and gene integration for Aspergillus niger. Bioresour Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 10.1016/j.biortech.2017.05.004. BB3_L 23_gRNAempty_cas9_BbsI; CRISPR plasmid with empty gRNA spacer to incorporate gRNA, Cas9 BB3_AMA_2.8_pUC-ORI_L_AC_hph
    pFREECas9, gRNA array (Synthetic)ptet, pRhamBAD Norholm A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z. Allows inducible expression of gRNAs and Cas9 for plasmid curing. pMAZ-SK
    pFREE_AmpCas9, gRNA array (Synthetic)ptet, pRhamBAD Norholm A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z. Allows inducible expression of gRNAs and Cas9 for plasmid curing. pMAZ-SK
    pFREE_CmCas9, gRNA array (Synthetic)ptet, pRhamBAD Norholm A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z. Allows inducible expression of gRNAs and Cas9 for plasmid curing. pMAZ-SK
    pFREE_ZeoCas9, gRNA array (Synthetic)ptet, pRhamBAD Norholm A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z. Allows inducible expression of gRNAs and Cas9 for plasmid curing. pMAZ-SK
    pFREE-RK2Cas9, gRNA array (Synthetic)ptet, pRhamBAD Norholm A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z. Allows inducible expression of gRNAs and Cas9 for plasmid curing as well as self-curing via a temperature sensitive RK2 replicon. pMAZ-SK
    pLQ-Pxyl/tet-cas9-Pj23119-sgRNACas9-Pxyltet-sgRNA-pj23119 (Other) Yang CRISPR/Cas9-based efficient genome editing in Staphylococcus aureus. Acta Biochim Biophys Sin (Shanghai). 2017 Sep 1;49(9):764-770. doi: 10.1093/abbs/gmx074. CRISPR-Cas9 based efficient genome editing in S. aureus unknown
    pET28a-Cas9-HisNLS-Cas9-NLS (Other)T7 Gao Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes. Nat Commun. 2017 Jan 18;8:14261. doi: 10.1038/ncomms14261. Purification of Cas9 protein pET28a (+)
    pJWV102-wtcas9spcas9sp (Synthetic)PczcD Kjos Chromosome segregation drives division site selection in Streptococcus pneumoniae. Proc Natl Acad Sci U S A. 2017 Jul 3. pii: 201620608. doi: 10.1073/pnas.1620608114. Plasmid containing wild-type cas9sp pJWV102
    pThermoCas9_ctrlCas9 from the type IIc CRISPR-Cas sytem of Geobacillus thermodenitrificans T12 strain (Other), ThermoCas9 single guide RNA expressing module (Other)B. smithii xylL promoter, B. coagulans DSM 1 pta promoter van der Oost Characterizing a thermostable Cas9 for bacterial genome editing and silencing. Nat Commun. 2017 Nov 21;8(1):1647. doi: 10.1038/s41467-017-01591-4. Expresses ThermoCas9 and its sgRNA module pNW33n
    SpyCas9(WT)SpyCas9 (Synthetic) Huang Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature. 2017 Jun 15;546(7658):436-439. doi: 10.1038/nature22377. Epub 2017 Apr 27. Express Streptococcus pyogenes Cas9 pGEX-6P-1
  • Tag / Fusion Protein
    • GST (N terminal on backbone)
    		•
    p6XHis_NLS-SaCas9CRISPR-associated protein Cas9/Csn1 [Staphylococcus aureus subsp. aureus] (Other)T7lac Tarleton Rapid, Selection-Free, High-Efficiency Genome Editing in Protozoan Parasites Using CRISPR-Cas9 Ribonucleoproteins. MBio. 2017 Nov 7;8(6). pii: e01788-17. doi: 10.1128/mBio.01788-17. Express SaCas9 in bacteria with a 6xHis tag for purification pET-32 EK/LIC
    pSHS207 - Bacterial expression plasmid for WT SpCas9SpCas9 (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for WT SpCas9 pCT10
    pJSC033 - Bacterial expression plasmid for SpCas9, REC2 FRET variantSpCas9 variant C80S/C574S/E60C/D273C (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9, REC2 FRET variant pCT10
    pJSC052 - Bacterial expression plasmid for SpCas9, REC3 FRET variantSpCas9 variant C80S/C574S/S701C/S960C (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9, REC3 FRET variant pCT10
    pSHS273 - Bacterial expression plasmid for SpCas9∆REC3 variantSpCas9 variant M1–N497,GGS,V713–D1368 (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9∆REC3 variant pCT10
    pJSC038 - Bacterial expression plasmid for SpCas9∆REC3, HNH FRET variantSpCas9 variant C80S/C574S/S355C/S867C/M1–N497,GGS,V713–D1368 (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9∆REC3, HNH FRET variant pCT10
    pJSC057 - Bacterial expression plasmid for SpCas9∆REC3, REC2 FRET variantSpCas9 variant C80S/C574S/E60C/D273C/M1–N497,GGS,V713–D1368 (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9∆REC3, REC2 FRET variant pCT10
    pSHS325 - Bacterial expression plasmid for SpCas9 REC3 domainSpCas9 variant K506–Q712 (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 REC3 domain pCT10
    pJSC176 - Bacterial expression plasmid for SpCas9 + Q926A variantSpCas9 variant Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 + Q926A variant pCT10
    pJSC119 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variantSpCas9 variant N497A/R661A/Q695A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variant pCT10
    pJSC120 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A, HNH FRET variantSpCas9 variant C80S/C574S/S355C/S867C/N497A/R661A/Q695A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A, HNH FRET variant pCT10
    pJSC111 - Bacterial expression plasmid for SpCas9-HF1 variantSpCas9 variant N497A/R661A/Q695A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9-HF1 variant pCT10
    pJSC115 - Bacterial expression plasmid for SpCas9-HF1, HNH FRET variantSpCas9 variant C80S/C574S/S355C/S867C/N497A/R661A/Q695A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9-HF1, HNH FRET variant pCT10
    pJSC129 - Bacterial expression plasmid for SpCas9-HF1, REC2 FRET variantSpCas9 variant C80S/C574S/E60C/D273C/N497A/R661A/Q695A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9-HF1, REC2 FRET variant pCT10
    pJSC131 - Bacterial expression plasmid for SpCas9-HF1, REC3 FRET variantSpCas9 variant C80S/C574S/S701C/S960C/N497A/R661A/Q695A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9-HF1, REC3 FRET variant pCT10
    pJSC090 - Bacterial expression plasmid for SpCas9 + K855A variantSpCas9 variant K855A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 + K855A variant pCT10
    pJSC077 - Bacterial expression plasmid for SpCas9 + K855A, HNH FRET variantSpCas9 variant C80S/C574S/S355C/S867C/K855A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 + K855A, HNH FRET variant pCT10
    pJSC114 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1) variantSpCas9 variant K848A/K1003A/R1060A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 eSpCas9(1.1) variant pCT10
    pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variantSpCas9 variant K848A/K1003A/R1060A/N497A/R661A/Q695A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant pCT10
    pJSC173 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9) variantSpCas9 variant N692A/M694A/Q695A/H698A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9) variant pCT10
    pJSC269 - Bacterial expression plasmid for SpCas9 Cluster 1 + Q926A variantSpCas9 variant N692A/M694A/Q695A/H698A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 1 + Q926A variant pCT10
    pJSC011 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), HNH FRET variantSpCas9 variant C80S/C574S/S355C/S867C/N692A/M694A/Q695A/H698A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), HNH FRET variant pCT10
    pJSC281 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC2 FRET variantSpCas9 variant C80S/C574S/E60C/D273C/N692A/M694A/Q695A/H698A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC2 FRET variant pCT10
    pJSC282 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC3 FRET variantSpCas9 variant C80S/C574S/S701C/S960C/N692A/M694A/Q695A/H698A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC3 FRET variant pCT10
    pJSC197 - Bacterial expression plasmid for SpCas9 Cluster 2 + Q926A variantSpCas9 variant G528A/V583A/E584A/D585A/N588A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 2 + Q926A variant pCT10
    pJSC228 - Bacterial expression plasmid for SpCas9 Cluster 2 variantSpCas9 variant G582A/V583A/E584A/D585A/N588A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 2 variant pCT10
    pJSC239 - Bacterial expression plasmid for SpCas9 Cluster 3 + Q926A variantSpCas9 variant T657A/G658A/W659A/R661A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 3 + Q926A variant pCT10
    pJSC236 - Bacterial expression plasmid for SpCas9 Cluster 4 + Q926A variantSpCas9 variant F491A/M495A/T496A/N497A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 4 + Q926A variant pCT10
    pJSC272 - Bacterial expression plasmid for SpCas9 Cluster 5 + Q926A variantSpCas9 variant K918A/V922A/R925A/Q926A (Other)T7 Doudna Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268. Bacterial expression plasmid for SpCas9 Cluster 5 + Q926A variant pCT10
    pEM-Cas9HF1-indRecA56Cas9HF1 (Other), recA56pTet, pTet Lynch Managing the SOS Response for Enhanced CRISPR-Cas-Based Recombineering in E. coli through Transient Inhibition of Host RecA Activity. ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174. Modified from pEM-Cas9HF1 (Addgene ID: 89961) to co-express inducible recA56 to block recA-mediated double-strand break repair. pEM-Cas9HF1
    AsCpf1-2NLSAsCpf1-6xHis-MBP-TEV (Synthetic)T7 Doudna CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing. Nat Commun. 2017 Dec 8;8(1):2024. doi: 10.1038/s41467-017-01836-2. bacterial expression of AsCpf1-2NLS pET
    LbCpf1-2NLSLbCpf1 (Synthetic)T7 Doudna CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing. Nat Commun. 2017 Dec 8;8(1):2024. doi: 10.1038/s41467-017-01836-2. bacterial expression of LbCpf1-2NLS pET
    pSBY1_FnCpf1cgFnCpf1 (Other)hsp60 Yang A CRISPR-Cpf1-Assisted Non-Homologous End Joining Genome Editing System of Mycobacterium smegmatis. Biotechnol J. 2018 Sep;13(9):e1700588. doi: 10.1002/biot.201700588. Epub 2018 Aug 6. CRISPR system used to genome editing in Mycobacterium smegmatis, expresses Fn CPf1 pMV261
    pJK02Upstream & Downstream pyrE deletion region (Other), tetR, Cas9 (Other), gRNA Sorg Using CRISPR-Cas9-mediated genome editing to generate C. difficile mutants defective in selenoproteins synthesis. Sci Rep. 2017 Nov 7;7(1):14672. doi: 10.1038/s41598-017-15236-5. Clostridium difficile CRISPR-cas9 mutagenesis plasmid traJ oriT pMTL84151
    pKM126Upstream & Downstream pyrE deletion region (Other), tetR, Cas9 (Other), gRNA Sorg Using CRISPR-Cas9-mediated genome editing to generate C. difficile mutants defective in selenoproteins synthesis. Sci Rep. 2017 Nov 7;7(1):14672. doi: 10.1038/s41598-017-15236-5. Clostridium difficile CRISPR-cas9 mutagenesis plasmid Tn916 oriT pJS116
    pCas9cas9 (Other), Lambda red genes (Other)J23105, araBAD Pfleger Genetic tools for reliable gene expression and recombineering in Pseudomonas putida. J Ind Microbiol Biotechnol. 2018 Jan 3. pii: 10.1007/s10295-017-2001-5. doi: 10.1007/s10295-017-2001-5. Recombineering plasmid with constitutively expressed cas9 and the araBAD promoter expressing αβγ pSEVA224
    p2T-CAG-KKHSaCas9-BlastRKKH SaCas9Chicken ß-Actin Sherwood Predictable and precise template-free CRISPR editing of pathogenic variants. Nature. 2018 Nov;563(7733):646-651. doi: 10.1038/s41586-018-0686-x. Epub 2018 Nov 7. Confers constitutive expression of KKH SaCas9. p2T-CAG-MCS-BlastR NEWENTRY
    pCAS9counterCas9, sgRNAprab17, prab17 Salipante Efficient and Scalable Precision Genome Editing in Staphylococcus aureus through Conditional Recombineering and CRISPR/Cas9-Mediated Counterselection. MBio. 2018 Feb 20;9(1). pii: mBio.00067-18. doi: 10.1128/mBio.00067-18. Expresses CAS9 and sgRNA for targeted counterselection in S. aureus pCN-50
    pxCas9CR4TetR and xCas9 (Synthetic) Reisch pCas9CR4 with point mutations from Cas9X for NG PAM sites (unpublished) PCas9CR4 with the xCas9 mutations allowing NG PAM sites p15A origin
    pRPaCas9Cas9 (Other) Horn Inducible high-efficiency CRISPR-Cas9-targeted gene editing and precision base editing in African trypanosomes. Sci Rep. 2018 May 21;8(1):7960. doi: 10.1038/s41598-018-26303-w. Expresses Cas9 in T. brucei (2T1) cells pUC
    pCas9-J23109Cas9 (Other) Zhang Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572. Plasmid constitutively expressing Cas9 protein N.A.
    peSpCas9-J23109eSpCas9 (Other) Zhang Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572. Plasmid constitutively expressing eSpCas9 protein N.A.
    pCasPA Ji CRISPR/Cas9-based Genome Editing in Pseudomonas aeruginosa and Cytidine Deaminase-Mediated Base Editing in Pseudomonas Species. iScience. 2018 Aug 31;6:222-231. doi: 10.1016/j.isci.2018.07.024. Epub 2018 Aug 1. Bacterial expression of Cas9 nuclease and λ-Red system in Pseudomonas aeruginosa unknown
    pNS20-SpCas9-SNAPCas9 (Other) Schwank Covalent linkage of the DNA repair template to the CRISPR-Cas9 nuclease enhances homology-directed repair. Elife. 2018 May 29;7. pii: 33761. doi: 10.7554/eLife.33761. Bacterial vector for expression of Snap-tagged Streptococcus pyogenes Cas9 pET His6 MBP N10 TEV LIC cloning vector (2C-T)
    pET-21a_3xNLS_SpCas9_protein_expression3xNLS_SpCas9 (Synthetic)T7 Wolfe Highly efficient therapeutic gene editing of human hematopoietic stem cells. Nat Med. 2019 May;25(5):776-783. doi: 10.1038/s41591-019-0401-y. Epub 2019 Mar 25. Protein expression plasmid for 3xNLS SpCas9 in pET-21a backbone pET-21 a
    pET-21a_2xNLS_LbCpf1_protein_expression2xNLS_LbCpf1 (Synthetic)T7 Wolfe Editing aberrant splice sites efficiently restores beta-globin expression in beta-thalassemia. Blood. 2019 Jan 31. pii: blood-2019-01-895094. doi: 10.1182/blood-2019-01-895094. Protein expression plasmid for 2xNLS LbCpf1 in pET-21a backbone pET-21 a
    pJL1-SpCas9S. pyogenes Cas9 (Other)T7 Jewett BioBits Health: Classroom Activities Exploring Engineering, Biology, and Human Health with Fluorescent Readouts. ACS Synth Biol. 2019 May 7. doi: 10.1021/acssynbio.8b00381. In vitro expression of S. pyogenes Cas9 from the T7 promoter pJL1
    pCasKP-aprCas9 (Other) Ji Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18. Bacterial expression of Cas9 nuclease and lambda-Red system in Klebsiella Pneumoniae Unknown
    pCasKP-hphCas9 (Other) Ji Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18. Bacterial expression of Cas9 nuclease and lambda-Red system in Klebsiella Pneumoniae Unknown
    pHSB04XCas9, promotor PslpA-guide-RNA, homologous arms of Lb_1019 (Other) Yang Development of a RecE/T-Assisted CRISPR-Cas9 Toolbox for Lactobacillus. Biotechnol J. 2019 Jul;14(7):e1800690. doi: 10.1002/biot.201800690. Epub 2019 May 20. Genome editing for Lactobacillus brevis ATCC367 pCas
    pHSP02Cas9, promotor P11-guide-RNA, homologous arms of Lp_0537 (Other) Yang Development of a RecE/T-Assisted CRISPR-Cas9 Toolbox for Lactobacillus. Biotechnol J. 2019 Jul;14(7):e1800690. doi: 10.1002/biot.201800690. Epub 2019 May 20. Genome editing for Lactobacillus plantarum WCSF1 pCas
    p15A_SpyCas9_CmRCas9 from S. pyogenes Noireaux An educational module to explore CRISPR technologies with a cell-free transcription-translation system (unpublished) Expresses S. pyogenes Cas9 p15A, Chloramphenicol resistance NEWENTRY
    pCas9-J23113Cas9 (Other) Zhang Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572. Expression of Cas9 with a weak promoter p15A origin
    Nme2Cas9_pMCSG7Nme2Cas9 (Other)T7 Sontheimer A Compact, High-Accuracy Cas9 with a Dinucleotide PAM for In Vivo Genome Editing. Mol Cell. 2018 Dec 18. pii: S1097-2765(18)31033-5. doi: 10.1016/j.molcel.2018.12.003. Nme2Cas9 bacterial expression plasmid pMCSG7
    NmeCas9_3XNLS_pMCSG7NmeCas9 (Other) Sontheimer NmeCas9 is an intrinsically high-fidelity genome editing platform (unpublished) Bacterial expression of wtNmeCas9 with 3X NLS pMCSG7
    pCas9CR4-VQRSpCas9-VQR (Synthetic)pTet Reisch pCas9CR4 variants for increased targeting space (unpublished) pCas9CR4 with VQR mutations for changing PAM site specificity to NGAN or NGNG from Kleinstiver et al. 2015 pdCas9-bacteria
    pCas9CR4-NGSpCas9-NG (Synthetic)pTet Reisch pCas9CR4 variants for increased targeting space (unpublished) pCas9CR4 with mutations to change PAM site specificity to NG from Nishimasu et al. pdCas9-bacteria
    pCas9CR5SpCas9 (Synthetic)pTet Reisch pCas9CR4 variants for increased targeting space (unpublished) pCas9CR4 with stop codon to prevent translation of ssrA degradation tag pdCas9-bacteria
    pCas9CR5-NGSpCas9-NG (Synthetic)pTet Reisch pCas9CR4 variants for increased targeting space (unpublished) pCas9CR5 with mutations to change PAM site specificity to NG from Nishimasu et al. pdCas9-bacteria
    pJZ002 Moore Refactoring the Cryptic Streptophenazine Biosynthetic Gene Cluster Unites Phenazine, Polyketide, and Nonribosomal Peptide Biochemistry. Cell Chem Biol. 2019 Feb 15. pii: S2451-9456(19)30038-8. doi: 10.1016/j.chembiol.2019.02.004. CRISPR-cas9 targeting in E. coli with ampicillin resistance N/A
    pSU-araC-Cas9Cas9 (Other)araBAD Yang Multigene editing in the Escherichia coli genome using the CRISPR-Cas9 system. Appl Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14. Multigene editing in the Escherichia coli genome using the CRISPR-Cas9 system p15A
    pET-21a_2xNLS_FnCpf1_MBP_protein_expression2xNLS_FnCpf1 (Synthetic)T7 Wolfe Enhanced Cas12a editing in mammalian cells and zebrafish. Nucleic Acids Res. 2019 Mar 20. pii: 5403491. doi: 10.1093/nar/gkz184. Protein expression plasmid for 2xNLS FnCpf1 in pET-21a backbone pET-21 a
    pUC-fFuCas9-HTBNLS-hphCas9 and hygromycin resist gene (Other)gpdA;trpC Ji CRISPR/Cas9-Based Genome Editing in the Filamentous Fungus Fusarium fujikuroi and Its Application in Strain Engineering for Gibberellic Acid Production. ACS Synth Biol. 2019 Feb 15;8(2):445-454. doi: 10.1021/acssynbio.8b00478. Epub 2019 Jan 23. Multigene editing in the Fusarium fujikuroi genome using the CRISPR-Cas9 system pUC57
    pEJS1026-pMCSG7-HpaCas9HpaCas9 (Other) Sontheimer Potent Cas9 Inhibition in Bacterial and Human Cells by AcrIIC4 and AcrIIC5 Anti-CRISPR Proteins. MBio. 2018 Dec 4;9(6). pii: mBio.02321-18. doi: 10.1128/mBio.02321-18. Expresses a 6X-His tagged type II-C Cas9 from H. parainfluenzae in bacterial cells pMCSG7
    pEJS1027-pMCSG7-SmuCas9SmuCas9 (Other) Sontheimer Potent Cas9 Inhibition in Bacterial and Human Cells by AcrIIC4 and AcrIIC5 Anti-CRISPR Proteins. MBio. 2018 Dec 4;9(6). pii: mBio.02321-18. doi: 10.1128/mBio.02321-18. Expresses a 6X-His tagged type II-C Cas9 from S. muelleri in bacterial cells pMCSG7
    pCasAb-apr Ji A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of OxyR. Cell Chem Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi: 10.1016/j.chembiol.2019.09.003. Bacterial expression of Cas9 nuclease and RecAb recombination system in Acinetobacter baumannii pMMB67EH
    pCpf1 Zhang Expanding the potential of CRISPR-Cpf1 based genome editing technology in the cyanobacterium Anabaena PCC 7120. ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437. Bacterial genome editing plasmid with kanamycin resistance marker Unknown
    pCpf1-sp Zhang Expanding the potential of CRISPR-Cpf1 based genome editing technology in the cyanobacterium Anabaena PCC 7120. ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437. Bacterial genome editing plasmid with spectinomycin resistance marker Unknown
    pCpf1b Zhang Expanding the potential of CRISPR-Cpf1 based genome editing technology in the cyanobacterium Anabaena PCC 7120. ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437. Bacterial genome editing plasmid with kanamycin resistance marker and using sacB as a counter-selection marker Unknown
    pCpf1b-sp Zhang Expanding the potential of CRISPR-Cpf1 based genome editing technology in the cyanobacterium Anabaena PCC 7120. ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437. Bacterial genome editing plasmid with spectinomycin resistance marker and using sacB as a counter-selection marker Unknown
    pLdSaCNgRNA and SaCas9 (Other) Matlashewski Single-Strand Annealing Plays a Major Role in Double-Strand DNA Break Repair following CRISPR-Cas9 Cleavage in Leishmania. mSphere. 2019 Aug 21;4(4). pii: 4/4/e00408-19. doi: 10.1128/mSphere.00408-19. Expresses Staphylococcus aureus Cas9 (SaCas9) and its gRNA in Leishmania pSP72
    pLPhygSaCas9Staphylococcus aureus Cas9 (Other) Matlashewski Single-Strand Annealing Plays a Major Role in Double-Strand DNA Break Repair following CRISPR-Cas9 Cleavage in Leishmania. mSphere. 2019 Aug 21;4(4). pii: 4/4/e00408-19. doi: 10.1128/mSphere.00408-19. Expresses Staphylococcus aureus Cas9 (SaCas9) in Leishmania pSP72 NEWENTRY
    PCV-Cas9PCV2 (Other), Cas9 (Other) Gordon Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template. Commun Biol. 2018 May 31;1:54. doi: 10.1038/s42003-018-0054-2. eCollection 2018. Expresses Cas9 with an N-terminal HUH-tag called PCV2 pTD68 ORF1 PCV2_gp1
    Cas9-PCVPCV2 (Other), Cas9 (Other) Gordon Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template. Commun Biol. 2018 May 31;1:54. doi: 10.1038/s42003-018-0054-2. eCollection 2018. Expresses Cas9 with a C-terminal HUH-tag called PCV2 in E. coli pTD68 ORF1 PCV2_gp1
    spCas9-NLS-MAVcas9 (Other)T7 Akavia Double-Stranded Biotinylated Donor Enhances Homology-Directed Repair in Combination with Cas9 Monoavidin in Mammalian Cells. CRISPR J. 2018 Dec;1:414-430. doi: 10.1089/crispr.2018.0045. SpCas9-NLS 17 amino acid linker to monoavidin (SpCas9-NLS-MAV) pEC-K-MBP
    pET-21a_FnCpf1_2xNLS_protein_expressionFnCpf1_2xNLS (Synthetic) Wolfe Enhanced Cas12a editing in mammalian cells and zebrafish. Nucleic Acids Res. 2019 Mar 20. pii: 5403491. doi: 10.1093/nar/gkz184. Protein expression plasmid for 2xNLS FnCpf1 in pET-21a backbone pET-21 a
    pFD115Ptet (Other), cas9 (Other), term PpflB sgRNA (BsaI) (Synthetic)Ptet Bikard Gene silencing with CRISPRi in bacteria and optimization of dCas9 expression levels. Methods. 2019 Aug 1. pii: S1046-2023(18)30497-3. doi: 10.1016/j.ymeth.2019.07.024. pFD115 carries cas9 controlled by an aTc-inducible Ptet promoter, a sgRNA controlled constitutively from PpflB S. aureus promoter to clone a guide between two BsaI sites and oriT on pLZ12 vector pLZ12
    pLQ-KO-tgt50Cas9-Pxyltet-sgRNA-Pspac-donor-tgt50 (Other) Yang CRISPR/Cas9-based efficient genome editing in Staphylococcus aureus. Acta Biochim Biophys Sin (Shanghai). 2017 Sep 1;49(9):764-770. doi: 10.1093/abbs/gmx074. CRISPR-Cas9 based efficient genome editing in S. aureus ColE1 origin, AmpR, Rep(+), cat resistance
    pIgnaviCas9IgnaviCas9 (Other)T7 promoter Quake Nucleic acid cleavage with a hyperthermophilic Cas9 from an uncultured Ignavibacterium. Proc Natl Acad Sci U S A. 2019 Oct 28. pii: 1904273116. doi: 10.1073/pnas.1904273116. Expresses IgnaviCas9 in E. coli pMJ806
    pKM197Upstream & Downstream pyrE deletion region (Other) Sorg Modified / Updated Clostridium difficile CRISPR-Cas9 genome editing plasmids (unpublished) pyrE-targeted CRISPR-Cas9 control plasmid. Cas9 expression is controlled by the xylR promoter and results in improved conjugation efficiency. Induction with xylose results in a pyrE mutant. pJS116
    pSCBE3-HFHF-BE3 (Other) Sun Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137 Expresses HF-BE3 CRISPR base editor and gRNA scaffold in Streptomyces pYH7

    Base Edit

    Catalytically dead dCas9 fused to a cytidine deaminase protein becomes a specific cytosine base editor that can alter DNA bases without inducing a DNA break. Cytosine base editors convert C->T (or G->A on the opposite strand) within a small editing window specified by the gRNA. Adenine base editors convert adenine to inosine, which is replaced by guanosine to create A->G (or T->C on the opposite strand) mutations.
    Plasmid Promoter PI Publication Hidden Extra Search Info
    pET42b-BE3T7 Liu Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery. Nat Commun. 2017 Jun 6;8:15790. doi: 10.1038/ncomms15790. Expresses BE3-NLS with an N-terminal His Tag (His6) for bacterial expression pET_42b
    pET42b-HF-BE3 Liu Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery. Nat Commun. 2017 Jun 6;8:15790. doi: 10.1038/ncomms15790. Expresses HF-BE3-NLS with an N-terminal His Tag (His6) for bacterial expression pET_42b
    pET28b-BE3∆UGIT7 Kim Genome-wide target specificities of CRISPR RNA-guided programmable deaminases. Nat Biotechnol. 2017 May;35(5):475-480. doi: 10.1038/nbt.3852. Epub 2017 Apr 10. The plasmid encoding the His6-rAPOBEC1-XTEN-nCas9 protein (BE3∆UGI) was generated by site-directed mutagenesis using pET28b-BE1 (Addgene plasmid #73018). pET28b
    pScI_dCas9-CDAlambda OR Kondo Deaminase-mediated multiplex genome editing in Escherichia coli. Nat Microbiol. 2018 Feb 5. pii: 10.1038/s41564-017-0102-6. doi: 10.1038/s41564-017-0102-6. Bacterial Target-AID vector pSC101
    pnCasPA-BEC Ji CRISPR/Cas9-based Genome Editing in Pseudomonas aeruginosa and Cytidine Deaminase-Mediated Base Editing in Pseudomonas Species. iScience. 2018 Aug 31;6:222-231. doi: 10.1016/j.isci.2018.07.024. Epub 2018 Aug 1. A cytidine deaminase-mediated base-editing plasmid in Pseudomonas species unknown
    pBECKP-km Ji Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18. A cytidine deaminase-mediated base-editing plasmid in Klebsiella Pneumoniae Unknown
    pBECKP-spe Ji Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18. ncas9-BEC, fusion protein of rAPOBEC1 and spCas9 nickase Unknown
    pET42b-A3A-PBE-ΔUGIT7 promoter Gao Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A. Nat Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261. To express the recombinant APOBEC3A in vitro pET42b
  • Tags / Fusion Proteins
    • 6*HIS (N terminal on insert)
    • NLS (C terminal on insert)
    		•
    pET42b-ABE7.10T7 Huang Genome-wide profiling of adenine base editor specificity by EndoV-seq. Nat Commun. 2019 Jan 8;10(1):67. doi: 10.1038/s41467-018-07988-z. Expresses ABE7.10-NLS with an N-terminal His Tag (His6) for bacterial expression pET42b
    pBECAb-apr Ji A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of OxyR. Cell Chem Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi: 10.1016/j.chembiol.2019.09.003. A cytidine deaminase-mediated base-editing plasmid in Acinetobacter baumannii pET28a and pWH1266
    pCRISPR-aBESTtipA, tipA Weber Highly efficient DSB-free base editing for streptomycetes with CRISPR-BEST. Proc Natl Acad Sci U S A. 2019 Oct 8;116(41):20366-20375. doi: 10.1073/pnas.1913493116. Epub 2019 Sep 23. Expression of a Streptomyces codon optimized Cas9n-tadA fusion protein in pGM1190. A to G base editor for actinomycetes pGM1190
    pSCBE2 Sun Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137 Expresses BE2 CRISPR base editor and gRNA scaffold in Streptomyces pYH7
    pSCBE3 Sun Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137 Expresses BE3 CRISPR base editor and gRNA scaffold in Streptomyces pYH7
    pSABEd Sun Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137 Expresses ABEd CRISPR base editor and gRNA scaffold in Streptomyces pYH7
    pSABEn Sun Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137 Expresses ABEn CRISPR base editor and gRNA scaffold in Streptomyces pYH7

    Nick

    CRISPR/Cas nickase mutants introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type Cas enzymes. To use a nickase mutant, you will need two gRNAs that target opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted off-target effects. Nickase mutants can also be used with a repair template to introduce specific edits via homology-directed repair (HDR).

    Plasmid Gene/Insert Promoter PI Publication Hidden Extra Search Info
    pMJ825Cas9T7 Doudna A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012 Jun 28. pEC-K-MBP
  • Tag / Fusion Protein
    • His6 (N terminal on backbone)
    		•
    pMJ826Cas9T7 Doudna A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012 Jun 28. pEC-K-MBP
  • Tag / Fusion Protein
    • His6 (N terminal on backbone)
    		•
    pMCSG7-D16A-NmeCas9pMCSG7-D16A-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 Nme cas9 nickase with a D16A mutation (RuvC domain) with T7 promoter, N-terminal His tag and TEV site pMCSG7
    pMCSG7-H588A-NmeCas9pMCSG7-H588A-NmeCas9 (Other)T7 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 Nme cas9 nickase with H588A mutation (HNH domain) with T7 promoter, N-terminal His tag and TEV site pMCSG7
    pNICKclos2.0Cas9 nickase (Other), sgRNA to xylR (Synthetic)Pthl, Pj23119 Yang CRISPR-based genome editing and expression control systems in Clostridium acetobutylicum and Clostridium beijerinckii. Biotechnol J. 2016 May 23. doi: 10.1002/biot.201600053. Genome editing for gene xylR (cbei-2385) in clostridium beijerinckii NCIMB 8052 pXY1
    pNICKclos1.0Cas9 nickase (Other), sGRNA to pyrEptb, j23119 Yang CRISPR-based genome editing and expression control systems in Clostridium acetobutylicum and Clostridium beijerinckii. Biotechnol J. 2016 May 23. doi: 10.1002/biot.201600053. Genome editing for gene pyrE (CAC-002) in Clostridium acetobutylicum ATCC 824 pIMP1-ptb
    pCas9(D10A)Cas9 D10A (Synthetic)pTeto Wang Targeted Large-Scale Deletion of Bacterial Genomes Using CRISPR-Nickases. ACS Synth Biol. 2015 Nov 20;4(11):1217-25. doi: 10.1021/acssynbio.5b00132. Epub 2015 Oct 25. Nicking Cas9 derived from pWT Cas9 (Addgene #44250). Ampicillin resistance marker, the tetracycline repressor (TetR), a ColE1 origin of replication and Streptococcus pyogenes Cas9 D10A pWTCas9
    pLCNICKCas9 nickase (Other), sgRNA, homology arms of LC2W_2179 Yang CRISPR-Cas9D10A Nickase-Assisted Genome Editing in Lactobacillus casei. Appl Environ Microbiol. 2017 Sep 1. pii: AEM.01259-17. doi: 10.1128/AEM.01259-17. Genome editing for Lactobacillus casei Lc2W pCas

    Activate

    Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest. If the plasmid that you choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator to your specific locus.

    Plasmid Gene/Insert Promoter PI Publication Hidden Extra Search Info
    pWJ66tracrRNA (Other), dcas9-w (Other), CRISPR array Marraffini Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic Acids Res. 2013 Jun 12. Same as pdCas9, but with the dCas9-w fusion (w is fused at the C-terminal end of dCas9) pACYC184
    pWJ68tracrRNA (Other), w-dcas9 (Other), CRISPR array Marraffini Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic Acids Res. 2013 Jun 12. Same as pdCas9, but with the w-dCas9 fusion (w is fused at the N-terminal end of dCas9) pACYC184
    pET-dCas9-VP64-6xHisdCas9-VP64 (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 dCas9-VP64-6xHis in bacterial cells pET29
    pET-deSpCas9-VP64-6xHisdead/inactive eSpCas9-NLS-3xFLAG-VP64 (Other)T7 Welker Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage. Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8. Expression of dead/inactive increased fidelity eSpCas9 (1.1)-VP64-6xHis in bacterial cells pET29
    pET-dSpCas9-HF1-VP64-6xHisdead/inactive SpCas9-HF1-NLS-3xFLAG-VP64 (Other)T7 Welker Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage. Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8. Expression of dead/inactive increased fidelity SpCas9-HF1-VP64-6xHis in bacterial cells pET29
    pET-dHeFSpCas9-VP64-6xHisdead/inactive HeFSpCas9-NLS-3xFLAG-VP64 (Other)T7 Welker Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage. Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8. Expression of dead/inactive increased fidelity HeFSpCas9-VP64-6xHis in bacterial cells pET29
    pFD116Ptet (Other), dcas9 (Other), PpflB sgRNA BsaI (Synthetic) Bikard Gene silencing with CRISPRi in bacteria and optimization of dCas9 expression levels. Methods. 2019 Aug 1. pii: S1046-2023(18)30497-3. doi: 10.1016/j.ymeth.2019.07.024. pFD116 carries dcas9 controlled by an aTc-inducible Ptet promoter, a sgRNA controlled constitutively from PpflB S. aureus promoter to clone a guide between two BsaI sites and oriT on pLZ12 vector pLZ12

    Interfere

    Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down gene expression by interfering with transcription. Design your gRNA to target your gene of interest’s promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-repressor to your specific locus.

    Plasmid Gene/Insert Promoter PI Publication Hidden Extra Search Info
    pMJ841Cas9 (Other)T7 Doudna A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012 Jun 28. pEC-K-MBP
  • Tags / Fusion Proteins
    • MBP (N terminal on backbone)
    • His6 (N terminal on backbone)
    		•
    pdCas9-bacteriadCas9 (bacteria) (Other)pLtetO-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 a catalytically inactive bacterial Cas9 (S. pyogenes) for bacterial gene knockdown p15A vector
    pdCas9tracrRNA (Other), dcas9 (Other), CRISPR array Marraffini Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic Acids Res. 2013 Jun 12. Expresses the tracrRNA, the dCas9 catalytic site mutant and a CRISPR array designed for the easy cloning of new spacers. pACYC184
    10xHis-MBP-TEV-S. pyogenes dCas9 M1C D10A C80S H840A C574SdCas9 M1C D10A C80S H840A C574S (Other) Doudna Programmable RNA recognition and cleavage by CRISPR/Cas9. Nature. 2014 Sep 28. doi: 10.1038/nature13769. dCas9 with single cysteine residue (M1C) for site-specific labelling pHMGWA
    pAN-PTet-dCas9dCas9 (Other)Ptet Voigt Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks. Mol Syst Biol. 2014 Nov 24;10:763. doi: 10.15252/msb.20145735. controls the expression of S. pyogenes dCas9 from a Tc‐inducible PTet promoter. unknown
    pCRISPathBrickConstitutive native promoters Koffas CRISPathBrick: Modular Combinatorial Assembly of Type II-A CRISPR Arrays for dCas9-Mediated Multiplex Transcriptional Repression in E. coli. ACS Synth Biol. 2015 Mar 30. E. coli vector for expression of S. pyogenes dCas9, tracrRNA, and nontargeting CRISPR array with BsaI site for inserting user-defined spacer-repeat bricks pdCas9-Marraffini (pACYC184)
    MSP712mammalian codon-optimized Streptococcus pyogenes dCas9 (D10A/H840A)-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 Sp-dCas9 & sgRNA (need to clone in spacer into BsaI sites): T7-humanSpdCas9(D10A/H840A)-T7-BsaIcassette-Sp-sgRNA pACYCDuet-1
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)
    		•
    pMM704dCas9, LacI, sgRNA Lu Programming a Human Commensal Bacterium, Bacteroides thetaiotaomicron, to Sense and Respond to Stimuli in the Murine Gut Microbiota Cell Systems, 2015 IPTG-inducible CRISPRi vector targeting BT1854, pNBU2 backbone, AmpR pExchange-tdk
    pMD19T-psba1-Ppsba2-dCas9-SpRdCas9 from S. pyogenes (Other)PpsbA2 Hudson Multiple Gene Repression in Cyanobacteria Using CRISPRi. ACS Synth Biol. 2015 Dec 28. Contains dCas9 from S. pyogenes under constitutive promoter. Suicide vector inserts into psba1 site of Synechocystis. Carries spectinomycin resist. Recommend E. coli Copy cutter for propogation. pMD19T simple
    pMD19T-psba1-TetR-PL22-dCas9-SpRdCas9 from S. pyogenes (Other)PL22 Hudson Multiple Gene Repression in Cyanobacteria Using CRISPRi. ACS Synth Biol. 2015 Dec 28. Contains dCas9 from S. pyogenes under aTc inducible promoter. Suicide vector inserts into psba1 site of Synechocystis. Carries spectinomycin resist. Recommend E. coli Copy cutter for propogation. pMD19T simple
    pdCas9-M-C4Repressor C4 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant C4. pdCas9
    pdCas9-M-3F2Repressor C4 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 3F2. pdCas9
    pdCas9-M-3H5Repressor 3H5 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 3H5. pdCas9
    pdCas9-M-1B6Repressor 1B6 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 1B6. pdCas9
    pdCas9-M-4F2Repressor 4F2 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 4F2. pdCas9
    pdCas9-M-5F5Repressor 5F5 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 5F5. pdCas9
    pdCas9-M-1D4Repressor 1D4 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 1D4. pdCas9
    pdCas9-M-4A6Repressor 4A6 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 4A6. pdCas9
    pdCas9-M-3A2Repressor 3A2 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 3A2. pdCas9
    pdCas9-M-1E4Repressor 1E4 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 1E4. pdCas9
    pdCas9-M-G6Repressor G6 (orthogonal T7-lac repressor) (Synthetic)Constitutive wild-type S. pyogenes promoter Koffas Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant G6. pdCas9
    pZ8-T_dCas9dcas9ptac Lu Corynebacterium glutamicum Metabolic Engineering with CRISPR Interference (CRISPRi). ACS Synth Biol. 2016 Feb 16. pZ8-1 plasmid carrying dcas9, driven by the IPTG-inducible Ptac promoter, KanR pZ8-1
    pZ8-P_dCas9dcas9Propionate inducible promoter (prp) Lu Corynebacterium glutamicum Metabolic Engineering with CRISPR Interference (CRISPRi). ACS Synth Biol. 2016 Feb 16. pZ8-1 plasmid carrying dcas9 driven by the propionate-inducible prpD2 promoter (PprpD2), KanR pZ8-Prp
    pJMP1dCas9 (Other)xylA Gross A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria. Cell. 2016 Jun 2;165(6):1493-506. doi: 10.1016/j.cell.2016.05.003. Epub 2016 May 26. Bacillus subtilis dCas9 expression vector; integrates into lacA/ganA pAX01
    pCas2AdCas9PEZ3 Pfleger CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007. dCas9 expressed from aTc-inducible promoter in acsA, A RBS: AGGAGA pACSA
    pCas2BdCas9PEZ3 Pfleger CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007. dCas9 expressed from aTc-inducible promoter in acsA, B RBS: TCGAGA pACSA
    pCas2CdCas9PEZ3 Pfleger CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007. dCas9 expressed from aTc-inducible promoter in acsA, C RBS: TGGACA pACSA
    pCas2DdCas9PEZ3 Pfleger CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007. dCas9 expressed from aTc-inducible promoter in acsA, D RBS: AGGACG pACSA
    pCas2EdCas9PEZ3 Pfleger CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007. dCas9 expressed from aTc-inducible promoter in acsA, E RBS: AGGGCG pACSA
    pCas2FdCas9PEZ3 Pfleger CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007. dCas9 expressed from aTc-inducible promoter in acsA, F RBS: TGGGCG pACSA
    pCas7dCas9c225 Pfleger CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007. dCas9 expressed from low constituve promoter, c225, in acsA pACSA
    pCas8dCas9none Pfleger CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007. dCas9 without a promoter in acsA pACSA
    pRH2502dcas9 (Other)uv15tetO Husson Investigating essential gene function in Mycobacterium tuberculosis using an efficient CRISPR interference system. Nucleic Acids Res. 2016 Jul 12. pii: gkw625. Expression of dcas9 D10A H840A from a TetR-regulated uvtetO promoter pTC-0X-1L
    pAW019-2dcas9 (Other)PxylA (B. megaterium) Chou Development of a CRISPR-Cas9 toolkit for comprehensive engineering of Bacillus subtilis. Appl Environ Microbiol. 2016 Jun 3. pii: AEM.01159-16. Inducible dCas9 integration vector for Bacillus subtilis pAX01 (ColE1)
    pUC18-mini-Tn7T-Lac-dCas9 (Ptac)S. pasteurianus dCas9 (Other) Prather A Robust CRISPR Interference Gene Repression System in Pseudomonas. J Bacteriol. 2018 Mar 12;200(7). pii: JB.00575-17. doi: 10.1128/JB.00575-17. Print 2018 Apr 1. Tn7 integrating plasmid with S. pasteurianus dCas9 expressed from the Ptac promoter for expression in P. putida and P. fluorescens pUC18
    pUC18-mini-Tn7T-Plac-dCas9 (Plac)S. pasteurianus dCas9 (Other) Prather A Robust CRISPR Interference Gene Repression System in Pseudomonas. J Bacteriol. 2018 Mar 12;200(7). pii: JB.00575-17. doi: 10.1128/JB.00575-17. Print 2018 Apr 1. Tn7 integrating plasmid with S. pasteurianus dCas9 expressed from the Plac promoter for expression in P. aeruginosa pUC18
    Biomolecular FeedBack (FB)pJ23100mut-dCas9-T- phtpG1-sgRNA (Other) Ellis Burden-driven feedback control of gene expression. Nat Methods. 2018 Mar 26. pii: nmeth.4635. doi: 10.1038/nmeth.4635. This construct encodes for two units. The first contains a constitutively expressed dCas9 protein. The second contains an sgRNA cassette under the control of the burden-responsive htpG1 promoter. pAZ16 ( Plasmid has got AmpR and p15A origin)
    pSET-dCas9dCas9 (Synthetic)ermE*p Lu CRISPR/dCas9-Mediated Multiplex Gene Repression in Streptomyces. Biotechnol J. 2018 Jun 3. doi: 10.1002/biot.201800121. Expression of the dCas9 gene in Streptomyces pSET152
    pSET-dCas9-actII-4-NT-S1dCas9 (Synthetic), sgRNAermE*p, J23119 Lu CRISPR/dCas9-Mediated Multiplex Gene Repression in Streptomyces. Biotechnol J. 2018 Jun 3. doi: 10.1002/biot.201800121. Repression of gene expression in Streptomyces by CRISPRi pSET152
    pCT310dCas9 (Other)T7 Tjian dCas9-targeted locus-specific protein isolation method identifies histone gene regulators. Proc Natl Acad Sci U S A. 2018 Mar 20;115(12):E2734-E2741. doi: 10.1073/pnas.1718844115. Epub 2018 Mar 5. Bacterial expression of 6His-dCas9-3XFlag pET302/NT-His
    pdCas9-J23111dCas9 (Other) Zhang Pooled CRISPR interference screening enables genome-scale functional genomics study in bacteria with superior performance. Nat Commun. 2018 Jun 26;9(1):2475. doi: 10.1038/s41467-018-04899-x. Plasmid constitutively expressing dCas9 protein N.A.
    pdCas9-J23109dCas9 (Other) Zhang Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572. Plasmid constitutively expressing dCas9 protein N.A.
    peSpdCas9-J23109eSpdCas9 (Other) Zhang Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572. Plasmid constitutively expressing eSpdCas9 protein N.A.
    pNS38-SadCas9-SNAPCas9 (Other) Schwank Covalent linkage of the DNA repair template to the CRISPR-Cas9 nuclease enhances homology-directed repair. Elife. 2018 May 29;7. pii: 33761. doi: 10.7554/eLife.33761. Bacterial vector for expression of Snap-tagged Staphylococcus aureus dCas9 pET His6 MBP N10 TEV LIC cloning vector (2C-T)
    PLJR962Sth1 sgRNA scaffold (Other), Sth1 dCas9 (Other), TetR (Other), L5 attP (Other), L5 Int (Other), KanR (Other) Fortune Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform. Nat Microbiol. 2017 Feb 6;2:16274. doi: 10.1038/nmicrobiol.2016.274. Sth1 dCas9 TetR and KanR L5 Int attP for M. smegmatis custom design
    PLJR965Sth1 sgRNA scaffold (Other), Sth1 dCas9 (Other), TetR (deleted TetON) MtbCO (Other), L5 attP (Other), L5 Int (Other), KanR (Other) Fortune Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform. Nat Microbiol. 2017 Feb 6;2:16274. doi: 10.1038/nmicrobiol.2016.274. Sth1 dCas9 TetR and KanR L5 Int attP for M. tuberculosis custom design

    RNA Targeting

    Type VI CRISPR systems, including the enzymes Cas13a/C2c2 and Cas13b, target RNA rather than DNA. In bacteria, once they have recognized and cleaved the target RNA sequence, they adopt an enzymatically active state and can bind and cleave additional RNAs regardless of homology to the crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which require that each DNA target have high sequence identity to the spacer sequence and contain a PAM sequence just downstream of the sequence to be cleaved. This non-specific cleavage is thought to activate programmed cell death or dormancy for phage-infected bacterial cells so as to limit the spread of infection throughout the entire population.

    Plasmid Promoter PI Publication Hidden Extra Search Info
    pZ003 (LshC2c2 locus) Zhang Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems. Mol Cell. 2015 Nov 5;60(3):385-97. doi: 10.1016/j.molcel.2015.10.008. Epub 2015 Oct 22. Expresses LshC2c2 full locus pACYC184
    pZ004 (LseC2c2 locus) Zhang Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems. Mol Cell. 2015 Nov 5;60(3):385-97. doi: 10.1016/j.molcel.2015.10.008. Epub 2015 Oct 22. Expresses LseC2c2 locus with one spacer in array pET28a
    pC001 - huLshC2C2-MBP for bacterial expressionT7 Zhang C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Jun 2. pii: aaf5573. Expresses human codon-optimized LshC2c2 for purification in E. coli BPV00356 pET21GG2-His(6)-MBP_Flag-Avi
    pC002 - LshC2C2 locus into pACYC184 Zhang C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Jun 2. pii: aaf5573. Endogenous LshC2c2 locus cloned into pACYC184 pACYC184
    pC003 - LshC2C2 locus into pACYC184 for spacer cloning Zhang C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Jun 2. pii: aaf5573. LshC2c2 locus in pACYC184 with BsaI sites for spacer cloning pACYC184
    p2CT-His-MBP-Lbu_C2c2_WT Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802. Bacterial expression of WT L. buccalis C2c2 CRISPR effector. p2CT
    p2CT-His-MBP-Lbu_C2c2_R472A_H477A Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802. Bacterial expression of L. buccalis C2c2 CRISPR effector (HEPN nuclease 1 inactive mutant). p2CT
    p2CT-His-MBP-Lbu_C2c2_R1048A_H1053A Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802. Bacterial expression of L. buccalis C2c2 CRISPR effector (HEPN nuclease 2 inactive mutant). p2CT
    p2CT-His-MBP-Lbu_C2c2_R472A_H477A_R1048A_H1053A Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802. Bacterial expression of L. buccalis C2c2 CRISPR effector (double HEPN nuclease 1 and 2 inactive mutant). p2CT
    p2CT-His-MBP-Lse_C2c2_WT Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802. Bacterial expression of WT L. seeligeri C2c2 CRISPR effector p2CT
    p2CT-His-MBP-Lsh_C2c2_WT Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802. Bacterial expression of WT L. shahii C2c2 CRISPR effector p2CT
    p2CT-His-MBP-Lbu_C2c2_R1079A Doudna Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802. Bacterial expression of L. buccalis C2c2 CRISPR effector (R1079A- pre-crRNA processing mutant) p2CT
    pBZCas13bLac Zhang Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5. Bacterial expression for bzCas13b, driven by the lac promoter, and DR-spacer-DR sequence driven by js23119. New spacer sequences can be cloned in between the DRs by digesting the plasmid with BsaI. pACYC184
    pBZCas13b-HEPNLac Zhang Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5. Bacterial expression for bzCas13b and crRNA with both HEPN domains mutated. New spacers can be cloned by digesting with BsaI. pACYC184
    pPbcas13bLac Zhang Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5. Bacterial expression for pbCas13b, driven by the lac promoter, and DR-spacer-DR sequence driven by js23119. New spacer sequences can be cloned in between the DRs by digesting the plasmid with BsaI. pACYC184
    pC013 - Twinstrep-SUMO-huLwCas13a Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321. Twinstrep-SUMO-huLwCas13a for recombinant protein bacterial expression. Insert is human codon optimized but expresses well in bacteria. pET
    p2CT-His-MBP-Lbu_C2c2_E299A Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lbu_C2c2_K310A Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lbu_C2c2_N314A Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lbu_C2c2_R1072A Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lbu_C2c2_D1078A Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lbu_C2c2_K1080A Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lbu_C2c2_K1082A Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lbu_C2c2_K1087A Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lwa_Cas13a_WT Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lne_Cas13a_WT Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Lba_Cas13a_WT Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Ere_Cas13a_WT Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Cam_Cas13a_WT Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Rca_Cas13a_WT Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Hhe_Cas13a_WT Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    p2CT-His-MBP-Ppr_Cas13a_WT Doudna RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. Bacterial expression for Cas13a p2CT
  • Tag / Fusion Protein
    • His6-MBP-TEV site (N terminal on backbone)
    		•
    pC009 LshCas13a locus with targeting spacer Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321. LshCas13a locus into pACYC184 with targeting spacer pACYC184
    pC010 LshCas13a locus with nontargeting spacer Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321. LshCas13a locus into pACYC184 with nontargeting spacer pACYC184
    pC011 LwCas13a locus with targeting spacer Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321. LwCas13a locus into pACYC184 with targeting spacer pACYC184
    pC012 LwCas13a locus with nontargeting spacer Zhang Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321. LwCas13a locus into pACYC184 with nontargeting spacer pACYC184
    pC018 - LshCas13a from Leptotrichia shahii Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses LshCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC019 - LwCas13a from Leptotrichia wadei Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses LwCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC020 - LseCas13a from Listeria seeligeri Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses LseCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC021 - LbmCas13a from Lachnospiraceae bacterium MA2020 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses LbmCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC022 - LbnCas13a from Lachnospiraceae bacterium NK4A179 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses LbnCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC023 - CaCas13a from [Clostridium] aminophilum DSM 10710 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses CaCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC024 - CgCas13a from Carnobacterium gallinarum DSM 4847 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses CgCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC025 - Cg2Cas13a from Carnobacterium gallinarum DSM 4847 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses Cg2Cas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC026 - PpCas13a from Paludibacter propionicigenes WB4 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses PpCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC027 - LweCas13a from Listeria weihenstephanensis FSL R9-0317 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses LweCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC028 - LbfCas13a from Listeriaceae bacterium FSL M6-0635 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses LbfCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC029 - Lw2Cas13a from Leptotrichia wadei F0279 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses Lw2Cas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC030 - RcsCas13a from Rhodobacter capsulatus SB 1003 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses RcsCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC031 - RcrCas13a from Rhodobacter capsulatus R121 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses RcrCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pC032 - RcdCas13a from Rhodobacter capsulatus DE442 Zhang RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049. Expresses RcdCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
    pDuBir-Lbu-dCas13a-avitagT7 O'Connell RNA Binding and HEPN-Nuclease Activation Are Decoupled in CRISPR-Cas13a. Cell Rep. 2018 Jul 24;24(4):1025-1036. doi: 10.1016/j.celrep.2018.06.105. Dual expression of Lbu-dCas13a with a C-terminal avi-tag and BirA to biotinylate Lbu-dCas13a pDuBir
    pET28a-MH6-EsCas13dlac Biotechnologies Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein. Mol Cell. 2018 Mar 9. pii: S1097-2765(18)30173-4. doi: 10.1016/j.molcel.2018.02.028. Expresses E. coli codon optimized EsCas13d in the pET28a backbone. pET-28a(+)
    pET28a-MH6-RspCas13d_RspCasWYL1lac Biotechnologies Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein. Mol Cell. 2018 Mar 9. pii: S1097-2765(18)30173-4. doi: 10.1016/j.molcel.2018.02.028. Expresses E. coli codon optimized RspCas13d and RspCasWYL1 in the pET28a backbone. pET-28a(+)
    pET28a-MH6-RspCas13dlac Biotechnologies Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein. Mol Cell. 2018 Mar 9. pii: S1097-2765(18)30173-4. doi: 10.1016/j.molcel.2018.02.028. Expresses E. coli codon optimized RspCas13d in the pET28a backbone. pET-28a(+)
    pC0057 BzoCas13 (B01) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses BzoCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0058 PinCas13 (B02) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses PinCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0059 PbuCas13 (B03) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses PbuCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0060 AspCas13 (B04) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses AspCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0061 PsmCas13 (B05) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses PsmCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0062 RanCas13 (B06) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses RanCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0063 PauCas13 (B07) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses PauCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0065 Pin2Cas13 (B09) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses Pin2Cas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0067 PguCas13 (B11) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses PguCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0068 PspCas13 (B12) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses PspCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0070 Pin3Cas13 (B15) His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses Pin3Cas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pC0072 LbuCas13a His6-TwinStrep-SUMO-BsaI Zhang Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15. Expresses LbuCas13a from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
    pLsCas13aGG Beisel Modular one-pot assembly of CRISPR arrays enables library generation and reveals factors influencing crRNA biogenesis. Nat Commun. 2019 Jul 3;10(1):2948. doi: 10.1038/s41467-019-10747-3. Backbone plasmid for generating CRISPR arrays for LsCas13a. Contains a GFP-dropout cassette and a direct repeat. pBAD18

    Purify

    A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA and its associated proteins, RNA, and DNA. The enCHIP system uses an anti-FLAG antibody to immunoprecipitate FLAG-tagged Cas9. Design your gRNA sequence to direct dCas9 to a specific locus, avoiding known transcription factor and other protein binding sites.

    Plasmid Gene/Insert Promoter PI Publication
    3xFLAG-dCas9/p-bacteria 3xFLAG-dCas9 pLtetO-1 Fujii Efficient isolation of specific genomic regions and identification of associated proteins by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR. Biochem Biophys Res Commun. 2013 Aug 11. pii: S0006-291X(13)01329-6. doi: 10.1016/j.bbrc.2013.08.013.

    Empty gRNA Expression Vectors

    Select a gRNA expression plasmid based on factors such as selectable marker or cloning method. When using CRISPR, you will need to express both a Cas protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein act as all-in-one vectors, but their function is often limited to a single category (cut, nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with a wide variety of Cas-containing plasmids.

    gRNA Plasmid Promoter Cloning
    Enzyme(s)    		 Validated In Resistance Co-expressed Cas9 Depositing lab
    Cas9 species = S. pyogenes (PAM = NGG)
    pCRISPR BsaI E. coli,
    S. pneumoniae     		Kanamycin none, need Cas9 plasmid Marraffini
    pCas9 BsaI E. coli,
    S. pneumoniae     		Chloramphenicol yes, cut Marraffini
    pgRNA-bacteria BBa_J23119 SpeI + HindIII Ampicillin none, need Cas9 plasmid Qi

    Do you have suggestions for other plasmids that should be added to this list?

    Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource!