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CRISPR header icon CRISPR/Cas Plasmids: Repress Gene Expression

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A catalytically inactive Cas9 (dCas9) or dCas9-repressor peptide fusion can be used to knock-down gene expression by interfering with transcription of the gene. Design your gRNA sequence to direct the dCas9 repressor to a specific genomic sequence. Potential target locations can include promoter regions, regulatory regions, and early coding regions. 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 repressor.



Mammalian

Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
pdCas9-humanizeddead Cas9 with 3X NLS (Homo sapiens)MSCV LTR promoterPuromycin 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. Expression of a catalytically inactive, human codon-optimized Cas9 under the control of Murine Stem Cell retroVirus LTR promoter for mammalian gene knockdown pMSCVpuro
pdCas9::BFP-humanizeddCas9 fused to BFP (Homo sapiens)MSCV LTR promoterPuromycin 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. Expression of a catalytically inactive, human codon-optimized Cas9-BFP fusion under the control of Murine Stem Cell retroVirus LTR promoter for mammalian gene knockdown pMSCVpuro
pHR-SFFV-dCas9-BFPdCas9-BFP fusion (Homo sapiens)SFFV Weissman CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi: 10.1016/j.cell.2013.06.044. Human expression vector containing SFFV promoter, dCas9 that is fused to 2x NLS and tagBFP pHR
pHR-SFFV-dCas9-BFP-KRABdCas9-BFP-KRAB fusion (Homo sapiens)SFFV Weissman CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi: 10.1016/j.cell.2013.06.044. Human expression vector containing SFFV promoter, dCas9 that is fused to 2x NLS, tagBFP and a KRAB domain pHR
pcDNA-dCas9dCas9 (Other)CMV Gersbach RNA-guided gene activation by CRISPR-Cas9-based transcription factors. Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2600. Expresses inactivated S. pyogenes dCas9 (D10A, H840A) in mammalian cells pcDNA3.1
Cas9m2Cas9m2 (Other) Church CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering. Nat Biotechnol. 2013 Aug 1. doi: 10.1038/nbt.2675. Cas9 D10A+H840A pcDNA3.3_TOPO
Cas9m3Cas9m3 (Other) Church CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering. Nat Biotechnol. 2013 Aug 1. doi: 10.1038/nbt.2675. Cas9 D10A+D839A+H840A pcDNA3.3_TOPO
Cas9m4Cas9m4 (Other) Church CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering. Nat Biotechnol. 2013 Aug 1. doi: 10.1038/nbt.2675. Cas9 D10A+D839A+H840A+N863A pcDNA3.3_TOPO
pAC84-pCR8-dCas9dCas9(D10A;H840A) (Homo sapiens) Jaenisch Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system. Cell Res. 2013 Aug 27. doi: 10.1038/cr.2013.122. dCas9 on Gateway donor vector pCR8/GW/TOPO. Note: This is not for expression. It has to be transferred to a gateway destination vector for expression pCR8/GW/TOPO
pHAGE TRE dCas9dCas9 (Other)TRENeomycin (select with G418) Wolfe Cas9 effector-mediated regulation of transcription and differentiation in human pluripotent stem cells. Development. 2014 Jan;141(1):219-23. doi: 10.1242/dev.103341. Tet-regulatable dCas9 lentiviral expression vector pHAGE
pHAGE TRE dCas9-KRABdCas9 (Other)TRENeomycin (select with G418) Wolfe Cas9 effector-mediated regulation of transcription and differentiation in human pluripotent stem cells. Development. 2014 Jan;141(1):219-23. doi: 10.1242/dev.103341. Tet-regulatable dCas9-KRAB lentiviral expression vector pHAGE
pHAGE EF1α dCas9-KRABdCas9 (Other)EF1alphaPuromycin Wolfe Cas9 effector-mediated regulation of transcription and differentiation in human pluripotent stem cells. Development. 2014 Jan;141(1):219-23. doi: 10.1242/dev.103341. Constitutive dCas9-KRAB lentiviral expression vector pHAGE
pSLQ1658-dCas9-EGFPdCas9 fuse to EGFP (Homo sapiens)MSCV LTR promoterPuromycin Qi Dynamic Imaging of Genomic Loci in Living Human Cells by an Optimized CRISPR/Cas System. Cell. 2013 Dec 19;155(7):1479-91. doi: 10.1016/j.cell.2013.12.001. Template for NLS-dCas9-NLS-EGFP fusion protein for CRISPR imaging (the recipient vector can be TetON 3G promoter system) pMSCVpuro
pLV hUbC-dCas9-T2A-GFPhumanized dead Cas9 T2A GFP (Other)hUbCZeocin Gersbach Multiplex CRISPR/Cas9-based genome engineering from a single lentiviral vector. Nucleic Acids Res. 2014 Aug 13. pii: gku749. Co-expresses human optimized S. pyogenes dCas9 and GFP FUGW
pcDNA3.1-CibN-dCas9CibN-dCas9 (Arabidopsis thaliana)CMVNeomycin (select with G418) Gersbach A light-inducible CRISPR-Cas9 system for control of endogenous gene activation. Nat Chem Biol. 2015 Feb 9. doi: 10.1038/nchembio.1753. Expresses CibN-dCas9 in mammalian cells pcDNA3.1
pcDNA3.1-dCas9-CibNdCas9-CibN (Arabidopsis thaliana)CMVNeomycin (select with G418) Gersbach A light-inducible CRISPR-Cas9 system for control of endogenous gene activation. Nat Chem Biol. 2015 Feb 9. doi: 10.1038/nchembio.1753. Expresses dCas9-CibN in mammalian cells pcDNA3.1
pcDNA3.1-CibN-dCas9-CibNdCas9-CibN (Arabidopsis thaliana)CMVNeomycin (select with G418) Gersbach A light-inducible CRISPR-Cas9 system for control of endogenous gene activation. Nat Chem Biol. 2015 Feb 9. doi: 10.1038/nchembio.1753. Expresses CibN-dCas9-CibN in mammalian cells pcDNA3.1
pHR-SFFV-KRAB-dCas9-P2A-mCherryKRAB-dCas9-P2A-mCherry fusion (Homo sapiens)SFFVmCherry Weissman Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014 Oct 9. 2nd Generation Lentiviral vector. Expresses an N-terminal KRAB-dCas9 fusion protein and mCherry pHR
pcDNA-dCas9-HAS.pyogenes dCas9 with c-terminal HA tag (Synthetic)CMV Gersbach Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers. Nat Biotechnol. 2015 May;33(5):510-7. doi: 10.1038/nbt.3199. Epub 2015 Apr 6. encodes c-terminal HA-tagged S. pyogenes dCas9 driven by CMV promoter pcDNA3.1
pJZC77sgRNA, COM-KRABU6, CMVMarked by mCherry Qi Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 10.1016/j.cell.2014.11.052. sgRNA (no RNA aptamer addition) with COM-KRAB effector for mammalian cells MP177_U6 (derived from pSico)
pJZC78sgRNA + 1x COM binding module, COM-KRABU6, CMVMarked by mCherry Qi Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 10.1016/j.cell.2014.11.052. sgRNA + 1x COM with COM-KRAB effector for mammalian cells MP177_U6 (derived from pSico)
pJZC73sgRNA, COM-KRABU6, CMVMarked by mCherry Qi Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 10.1016/j.cell.2014.11.052. sgRNA (no RNA aptamer addition) with COM-KRAB effector for mammalian cells MP177_U6 (derived from pSico)
pJZC74sgRNA + 1x COM binding module, COM-KRABU6, CMVMarked by mCherry Qi Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 10.1016/j.cell.2014.11.052. sgRNA + 1x COM with COM-KRAB effector for mammalian cells MP177_U6 (derived from pSico)
pX330A_dCas9-1x2humanized S. pyogenes dCas9 (Other)CBh Yamamoto Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. BMC Biotechnol. 2015 May 22;15(1):33. Expresses dCas9 and gRNA pUC ori vector
pX330A_dCas9-1x3humanized S. pyogenes dCas9 (Other)CBh Yamamoto Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. BMC Biotechnol. 2015 May 22;15(1):33. Expresses dCas9 and gRNA pUC ori vector
pX330A_dCas9-1x5humanized S. pyogenes dCas9 (Other)CBh Yamamoto Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. BMC Biotechnol. 2015 May 22;15(1):33. Expresses dCas9 and gRNA pUC ori vector
pX330A_dCas9-1x6humanized S. pyogenes dCas9 (Other)CBh Yamamoto Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. BMC Biotechnol. 2015 May 22;15(1):33. Expresses dCas9 and gRNA pUC ori vector
pEF_dCas9dCas9 (Other)human EF1[alpha] Rinn Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display. Nat Methods. 2015 Jul;12(7):664-70. doi: 10.1038/nmeth.3433. Epub 2015 Jun 1. Transient expression of Sp-dCas9 in mammalian cells, under an EF1-alpha promoter. pNEB193
  • Tag / Fusion Protein
    • 3xFLAG (C terminal on insert)
    		•
    pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-Purohumanized dCas9-KRAB T2A Puro (Other), sgRNAPuromycin Gersbach Highly specific epigenome editing by CRISPR-Cas9 repressors for silencing of distal regulatory elements. Nat Methods. 2015 Dec;12(12):1143-9. doi: 10.1038/nmeth.3630. Epub 2015 Oct 26. Express sgRNA and dCas9-KRAB from lentiviral vector FUGW
    pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-GFPhumanized dCas9-KRAB T2A GFP (Other), sgRNA Gersbach Highly specific epigenome editing by CRISPR-Cas9 repressors for silencing of distal regulatory elements. Nat Methods. 2015 Dec;12(12):1143-9. doi: 10.1038/nmeth.3630. Epub 2015 Oct 26. Express sgRNA and dCas9-KRAB from lentiviral vector FUGW
    pCR1003T7pr_10xHis-MBP-TEV-SPydCas9 (D10A, H840A) Corn Enhancing homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Nat Biotechnol. 2016 Jan 20. doi: 10.1038/nbt.3481. Express Streptococcus pyogenes dCas9 (D10A, H840A) SPynCas9
    pCR1056T7pr_6xHis-MBP-TEV-SPydCas9 (D10A,H840A)-2xNLS Corn Enhancing homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Nat Biotechnol. 2016 Jan 20. doi: 10.1038/nbt.3481. Express Streptococcus pyogenes dCas9 (D10A, H840A) carrying two C-terminal SV40 NLS SPynCas9
    pAC1445-pmax-dCas9dCas9 (Synthetic)CAGGS + chim intron Cheng pAC1445 (unpublished) dCas9 expressed in pmax vector pAC90-pmax-DEST
    pAAVS1-NDi-CRISPRi (Gen1)dCas9-KRAB-P2A-mCherry (Synthetic), rtTA (Synthetic)TRE3G, CAGNeomycin (select with G418) Conklin CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs. Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi: 10.1016/j.stem.2016.01.022. Epub 2016 Mar 10. Dox-inducible CRISPR interference (CRISPRi) knock in construct into the AAVS1 locus with mCherry marker pAAVS1
    pAAVS1-NDi-CRISPRi (Gen2)dCas9-KRAB (Synthetic), rtTA (Synthetic)TRE3G, CAGNeomycin (select with G418) Conklin CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs. Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi: 10.1016/j.stem.2016.01.022. Epub 2016 Mar 10. Dox-inducible CRISPR interference (CRISPRi) knock in construct into the AAVS1 locus pAAVS1
    pAAVS1-NC-CRISPRi (Gen3)dCas9-KRAB (Synthetic)CAGNeomycin (select with G418) Conklin CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs. Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi: 10.1016/j.stem.2016.01.022. Epub 2016 Mar 10. Constitutive CRISPR interference (CRISPRi) knock in construct into the AAVS1 locus pAAVS1
    pHAGE-TO-dCas9Sp dCas9 (Synthetic)CMV-TO Pederson Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526. dCas9 pHAGE-DEST
    CSII-U6-gRNA-CBh-3xFLAG-PA-dCas9-P2A-PurosgRNA and dCas9 from pX330 (Other)U6 for sgRNA and CBh for dCas9Puromycin Kimura Dr. Sekita CRISPR/Cas9 (unpublished) Lentivirus vector to express guideRNA and dCas9 with puro resistant gene CSII
    pLV-dCas9-KRAB-PGK-HygRhumanized dead Cas9 KRAB (Other), aminoglycoside phosphotransferase from E. coli (Other)Human Ubiquitin C Promoter, mouse phosphoglycerate kinase 1 promoterHygromycin Gersbach CRISPR-Cas9 epigenome editing enables high-throughput screening for functional regulatory elements in the human genome. Nat Biotechnol. 2017 Apr 3. doi: 10.1038/nbt.3853. Lentiviral Sp dCas9-KRAB fusion with Hygromycin B resistance cassette. FUGW
    pSLQ2818 pPB: CAG-PYL1-KRAB-IRES-Puro-WPRE-SV40PA PGK-ABI-tagBFP-SpdCas9ABI-tagBFP-Sp dCas9 (Arabidopsis thaliana), PYL1-KRAB (Arabidopsis thaliana)PGK, CAGPuromycin Qi Complex transcriptional modulation with orthogonal and inducible dCas9 regulators. Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042. Expresses ABA-inducible KRAB-Sp dCas9 PB
    pSLQ2813 pPB: CAG-GID1-KRAB-IRES-Puro-WPRE PGK-ABI-tagBFP-SpdCas9GAI-tagBFP-Sp dCas9 (Arabidopsis thaliana), GID1-KRAB (Arabidopsis thaliana)PGK, CAGPuromycin Qi Complex transcriptional modulation with orthogonal and inducible dCas9 regulators. Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042. Expresses GA-inducible KRAB-Sp dCas9 PB
    pSLQ2815 pPB: CAG-Puro-WPRE PGK-KRAB-tagBFP-SpdCas9KRAB-tagBFP-Sp dCas9 (Synthetic), PuroPGK, CAGPuromycin Qi Complex transcriptional modulation with orthogonal and inducible dCas9 regulators. Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042. Expresses direct fusion KRAB-Sp dCas9 PB
    pGH125_dCas9-BlastdCas9 and Blasticidin resistanceBlasticidin Bassik Directed evolution using dCas9-targeted somatic hypermutation in mammalian cells. Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4038. lentiviral expression vector for dCas9 with Blasticidin selectable marker Addgene #61425
    TRE-KRAB-dCas9-IRES-BFPKRAB-dCas9-IRES-BFP (Synthetic)TRE3GBFP Lander Systematic mapping of functional enhancer-promoter connections with CRISPR interference. Science. 2016 Sep 29. pii: aag2445. Lentiviral vector expressing a KRAB-dCas9 fusion protein and BFP pHR
    pMH0001dCas9 (Other)SFFVBFP fluorescence Weissman A Multiplexed Single-Cell CRISPR Screening Platform Enables Systematic Dissection of the Unfolded Protein Response. Cell. 2016 Dec 15;167(7):1867-1882.e21. doi: 10.1016/j.cell.2016.11.048. UCOE-SFFV-dCas9-BFP-KRAB pHR-SFFV-dCas9-BFP-KRAB
    Lenti-dCas9-KRAB-blastdCas9 (Synthetic)Blasticidin Hon Multiplexed Engineering and Analysis of Combinatorial Enhancer Activity in Single Cells. Mol Cell. 2017 Apr 20;66(2):285-299.e5. doi: 10.1016/j.molcel.2017.03.007. Epub 2017 Apr 13. Plasmid expression dCas9 protein in fusion with KRAB domain plenti
    pLX_311-KRAB-dCas9dCas9 (Other)EF1aBlasticidin Hahn Complementary information derived from CRISPR Cas9 mediated gene deletion and suppression. Nat Commun. 2017 May 23;8:15403. doi: 10.1038/ncomms15403. for CRISPRi, lentiviral expression of KRAB-dCas9 and BlastR. Also called pXPR_121 pXPR
  • Tag / Fusion Protein
    • KRAB (N terminal on insert)
    		•


    Bacteria

    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
    DS-SPcasN-Cas9, nuclease-null (Other), tracrRNA precursor (Other)proC, tracdrRNA promoter Church Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. Bacterial nuclease-null SP Cas9 expression cloDF13-aadA
    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
    pET302-6His-dCas9-HaloHis6-dCas9-Halo (Synthetic) Lionnet CASFISH: CRISPR/Cas9-mediated in situ labeling of genomic loci in fixed cells. Proc Natl Acad Sci U S A. 2015 Sep 22;112(38):11870-5. doi: 10.1073/pnas.1515692112. Epub 2015 Aug 31. Encodes a Halo-labeled fusion of nuclease deficient Cas9 pET302
    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
    pdCASclosdCas9 (Other), sgRNA to spo0Aptb, 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. Transcriptional repression for gene Spo0A (CAC-2011) in Clostridium acetobutylicum ATCC 824 pIMP1-ptb
    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)
    Jun lab tunable CRISPRi strain Jun tCRISPRi: tunable and reversible, one-step control of gene expression. Sci Rep. 2016 Dec 20;6:39076. doi: 10.1038/srep39076. Strain SJ_XTL219 Genotype: MG1655 PBAD_dcas9 ΔlacI ΔaraE araFGH<>spec lacY A177C, galM<pBBa-J23119-tet-sacB-handle-term strain: SJ_XTL219


    Yeast

    Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    pTDH3-dCas9dCas9TDH3LEU2 Weissman CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi: 10.1016/j.cell.2013.06.044. Yeast CEN/ARS vector (Leu2) that contains dCas9 fused to NLS controlled by TDH3 promoter pJED103
    pTDH3-dCas9-Mxi1dCas9-Mxi1TDH3LEU2 Weissman CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi: 10.1016/j.cell.2013.06.044. Yeast CEN/ARS vector (Leu2) that contains dCas9 fused to NLS and Mxi1 domain controlled by TDH3 promoter pJED103
    pJZC518dCas9 (Other)pTdh3LEU2 Qi Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 10.1016/j.cell.2014.11.052. Yeast dCas9 expression plasmid pNH605
    pJZC572sgRNA + 1x com RNA binding moduleSNR52URA3 Qi Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 10.1016/j.cell.2014.11.052. sgRNA with 1x com for yeast cells pRS416
    pTPGI_dCas9Yeast-optimized dCas9 (Saccharomyces cerevisiae)pTPGITRP1 Lu Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas. ACS Synth Biol. 2013 Sep 11. encodes yeast-optimized dCas9 synthetic transcription factor pRS304
    pRS416-dCas9-Mxi1 + TetR + pRPR1(TetO)-NotI-gRNAdCas9-Mxi1 (Synthetic), Tet Repressor (Other), Structural gRNA for S pyogenes (Synthetic)pTef1, pGPM1, pRPR1(TetO)URA3 Davis Quantitative CRISPR interference screens in yeast identify chemical-genetic interactions and new rules for guide RNA design. Genome Biol. 2016 Mar 8;17(1):45. doi: 10.1186/s13059-016-0900-9. pRS416 Ura marked Cen/Ars plasmid with dCas9-Mxi1 under Tef1 promoter, and tet-incucibile RPR1 promoter with NotI cloning site adjacent to gRNA pRS416


    Plant

    Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    pBUN6I11dCas9-KRAB (Synthetic), gRNA scaffold (Synthetic)Ubi1p, OsU3pBar Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses dCas9-KRAB, gRNA scaffold for insertion of target sequence (OsU3 promoter), Bar resistance pGreen-like binary vector
    pHSN6I01dCas9-KRAB (Synthetic), gRNA scaffold (Synthetic)2×35Sp, AtU6-26pHygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses dCas9-KRAB, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Hyg resistance pGreen-like binary vector
    pEGB 35S:dCas9:Tnos (GB1191)dCas9 (Other)35S Orzaez GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. Plant Physiol. 2013 Jul;162(3):1618-31. Transcriptional unit for human codon optimized with mutated (D10A, H840A) and inactivated catalytic domains Cas9 protein plant expression driven by the 35S promoter pDGB3alpha2
    pYPQ153pco-dCas9-3X(SRDX) (Plant codon-optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with pco-dCas9-3X(SRDX) pYPQ185-linker1
    pdCas9 (GB1079)Cas9 coding region with mutated (D10A, H840A) and inactivated catalytic domains (human codon optimised) (Other) Orzaez A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. eCollection 2016. Provides the human codon optimized CDS of Cas9 protein with mutated (D10A, H840A) and inactivated catalytic domains as a level 0 GoldenBraid part for C-terminal fusions pUPD
    pEGB 35s:dCas:BRD:tNos (GB1172)dCas9:BRD (Synthetic)35S Orzaez A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. eCollection 2016. Transcriptional unit of (human codon optimized) inactivated Cas9 fused to the BRD Transcriptional Repressor pDGB3alpha2


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