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CRISPR Plasmids: Yeast

Browse CRISPR Plasmids

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CRISPR Resources

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

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 Selectable Marker PI Publication Hidden Extra Search Info
p414-TEF1p-Cas9-CYC1tHuman Optimized S. pyogenes Cas9 (Homo sapiens)TEF1 promoterTRP1 Church Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Nucleic Acids Res p414
p415-GalL-Cas9-CYC1tHuman Optimized S. pyogenes Cas9 (Homo sapiens)GalLLEU2 Church Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Nucleic Acids Res p415
pACT2-CAS9cCas9c (Synthetic)yeast ADH1 promoterLEU2 Zhao Self-processing of ribozyme-flanked RNAs into guide RNAs in vitro and in vivo for CRISPR-mediated genome editing. J Integr Plant Biol. 2013 Dec 30. doi: 10.1111/jipb.12152. Express Eukaryotic-codon-optimized Cas9c gene in yeast under ADH1 promoter for genome editing. SV40 NLS is fused to the C terminal. GAL4 region from pACT2 is removed. pACT2
pMZ222Cas9 (Other)adh1LEU2 Zaratiegui Implementation of the CRISPR-Cas9 system in fission yeast. Nat Commun. 2014 Oct 29;5:5344. doi: 10.1038/ncomms6344. Constitutive expression of humanized Cas9 pART1
pMZ374Cas9 (Other), rrk1:sgRNA (Other)adh1, rrk1ura4 Zaratiegui Implementation of the CRISPR-Cas9 system in fission yeast. Nat Commun. 2014 Oct 29;5:5344. doi: 10.1038/ncomms6344. Combination adh1:cas9/rrk1:sgRNA for CRISPR genome editing in fission yeast: Empty sgRNA target (CspCI placeholder) (see comments). pMZ283
pCTiCas9 (Other), tracrRNA (Other)TEF1p, RPR1pLEU2 Zhao Homology-Integrated CRISPR-Cas (HI-CRISPR) System for One-Step Multigene Disruption in Saccharomyces cerevisiae. ACS Synth Biol. 2014 Sep 19. Plasmid encoding iCas9 and tracrRNA on pRS415 backbone pRS415
pCRCTiCas9 (Other), tracrRNA (Other)TEF1p, RPR1pURA3 Zhao Homology-Integrated CRISPR-Cas (HI-CRISPR) System for One-Step Multigene Disruption in Saccharomyces cerevisiae. ACS Synth Biol. 2014 Sep 19. Plasmid encoding iCas9, tracrRNA and crRNAs pRS426
Cas9-NATHuman-optimized S. pyogenes Cas9 with Clonnat marker gene expression cassetteNourseothricin(clonNat) Jin Construction of a quadruple auxotrophic mutant of an industrial polyploid saccharomyces cerevisiae strain by using RNA-guided Cas9 nuclease. Appl Environ Microbiol. 2014 Dec;80(24):7694-701. doi: 10.1128/AEM.02310-14. Epub 2014 Oct 3. Cas9 expression cassette carried by a yeast single-copy episome plasmid pRS414-TEF1p-Cas9-CYC1t
pML104Cas9 (Saccharomyces cerevisiae), single guide RNA expression cassette (Saccharomyces cerevisiae)pTDH3, pSNR52URA3 Wyrick New vectors for simple and streamlined CRISPR-Cas9 genome editing in Saccharomyces cerevisiae. Yeast. 2015 Dec;32(12):711-20. doi: 10.1002/yea.3098. Epub 2015 Sep 21. Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains URA3 marker for yeast transformation pRSII426
pML107Cas9 (Saccharomyces cerevisiae), single guide RNA expression cassette (Saccharomyces cerevisiae)pTDH3 (aka GAP promoter), pSNR52LEU2 Wyrick New vectors for simple and streamlined CRISPR-Cas9 genome editing in Saccharomyces cerevisiae. Yeast. 2015 Dec;32(12):711-20. doi: 10.1002/yea.3098. Epub 2015 Sep 21. Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains LEU2 marker for yeast transformation pRSII425
pJH001Cas9 (Saccharomyces cerevisiae), EmptyLEU2 Wyrick New vectors for simple and streamlined CRISPR-Cas9 genome editing in Saccharomyces cerevisiae. Yeast. 2015 Dec;32(12):711-20. doi: 10.1002/yea.3098. Epub 2015 Sep 21. High copy Cas9 expression vector with LEU2 marker for yeast transformation pRSII425
p415-PtrpC-Cas9-TtrpC-CYC1thumanized Cas9 (Homo sapiens)LEU2 Hong Efficient gene editing in Neurospora crassa with CRISPR technology Fungal Biology and Biotechnology 2015, 2:4 Express humanized Cas9 under the control of trpC promoter and terminator from A. nidulans. pRS415
pCRISPRylCodon optimized Cas9 from S. pyogenes (Synthetic), sgRNA expression cassette (Synthetic)UAS1B8-TEF(136), SCR1'-tRNALEU2 Wheeldon Synthetic RNA polymerase III promoters facilitate high efficiency CRISPR-Cas9 mediated genome editing in Yarrowia lipolytica. ACS Synth Biol. 2015 Dec 29. CRISPR/Cas9 vector for Yarrowia lipolytica, with AvrII site for sgRNA insertion pUC19
pMZ376Cas9 (Other), rrk1:sgRNA (Other)adh1, rrk1Neomycin (select with G418) Zaratiegui sgRNA/Cas9 (unpublished) Combined sgRNA/Cas9 vector with KanMX marker pMZ374
pMZ377Cas9 (Other), rrk1:sgRNA (Other)adh1, rrk1LEU2 Zaratiegui sgRNA/Cas9 (unpublished) Combined sgRNA/Cas9 vector with LEU2 marker pMZ374
pMZ379Cas9 (Other), rrk1:sgRNA (Other)adh1, rrk1nourseothricin Zaratiegui sgRNA/Cas9 (unpublished) Combined sgRNA/Cas9 vector with NatMX marker pMZ374
pCfB2312 (TEF1p-Cas9-CYC1t_kanMX)Cas9 (Other)Gentamicin Borodina EasyClone-MarkerFree: A vector toolkit for marker-less integration of genes into Saccharomyces cerevisiae via CRISPR-Cas9. Biotechnol J. 2016 Aug;11(8):1110-7. doi: 10.1002/biot.201600147. Epub 2016 Jun 23. Expresses Cas9 protein, under the control of the TEF1 promoter, and CYC terminator. KanMX resistance pRS414
pRS416gT-GalL-Cas9Cas9 (Synthetic), guide RNA (gRNA) (Synthetic), Tetracycline Repressor (Other)GalL, RPR1 promoter with TetO site, GPM1 promoterURA3 Davis Distinct patterns of Cas9 mismatch tolerance in vitro and in vivo. Nucleic Acids Res. 2016 May 19. pii: gkw417. pRS416 (URA) + RPR1(TetO) promoter with gRNA locus + Tetracycline Repressor + GalL promoter driven Cas9 for yeast expression pRS416
pRCC-KCas9 (Other), empty gRNA cassette (Synthetic)ROX3, SNP52pNeomycin (select with G418) Boles Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. Expression of Cas9 and gRNA cassette in S. cerevisiae; Kan Resistance pRS42K
  • Tag / Fusion Protein
    • NTS (C terminal on backbone)
    		•
    pRCC-NCas9 (Other), empty gRNA cassette (Synthetic)ROX3, SNP52pNourseothricin Boles Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. Expression of Cas9 and gRNA cassette in S. cerevisiae; Nourseothricin Resistance pRS42N
  • Tag / Fusion Protein
    • NTS (C terminal on backbone)
    		•
    pDuRCC-KCas9 (Other), empty gRNA cassette (Synthetic), empty gRNA cassette (Synthetic)ROX3, SNP52p, SNP52pNeomycin (select with G418) Boles Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. Expression of Cas9 and two gRNA cassettes in S. cerevisiae; Kan Resistance pRS42K
  • Tag / Fusion Protein
    • NTS (C terminal on backbone)
    		•
    pDuRCC-NCas9 (Other), empty gRNA cassette (Synthetic), empty gRNA cassette (Synthetic)ROX3, SNP52p, SNP52pNourseothricin Boles Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. Expression of Cas9 and two gRNA cassettes in S. cerevisiae; Nourseothricin Resistance pRS42N
  • Tag / Fusion Protein
    • NTS (C terminal on backbone)
    		•
    pML104-HygMx4HphMX4 (Other)Hygromycin Wittkopp Patricia Wittkopp yeast CRISPR plasmids (unpublished) Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains HphMx4 marker for yeast transformation. pRSII426
    pML104-KanMx4KanMX4 (Other)Neomycin (select with G418) Wittkopp Patricia Wittkopp yeast CRISPR plasmids (unpublished) Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains KanMx4 marker for yeast transformation. pRSII426
    pML104-NatMx3NatMx3 (Other)nourseothricin (nat) Wittkopp Patricia Wittkopp yeast CRISPR plasmids (unpublished) Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains NatMX3 marker for yeast transformation. pRSII426
    pCfB2312 (TEF1p-Cas9-CYC1t_kanMX)Cas9 (Other)G418 Borodina CRISPR–Cas system enables fast and simple genome editing of industrial Saccharomyces cerevisiae strains Metab Eng Commun 2015 Cas9-carrying vector with kanMX marker p414-TEF1p-Cas9-CYC1t
    pCRISPRyl_AXPCas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at AXP locus in Y. lipolytica pUC19
    pCRISPRyl_XPR2Cas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at XPR2 locus in Y. lipolytica pUC19
    pCRISPRyl_A08Cas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at A08 locus in Y. lipolytica pUC19
    pCRISPRyl_D17Cas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at D17 locus in Y. lipolytica pUC19
    pCRISPRyl_MFE1Cas9UAS1B8-TEF(136)LEU2 Wheeldon Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. ACS Synth Biol. 2016 Dec 19. Introduces DSB at MFE1 locus in Y. lipolytica pUC19
    pJK-caCas9-NatMX-Neut5LNourseothricin Church A CRISPR-Cas9-based gene drive platform for genetic interaction analysis in Candida albicans. Nat Microbiol. 2018 Jan;3(1):73-82. doi: 10.1038/s41564-017-0043-0. Epub 2017 Oct 23. caCas9 integrating vector into the Neut5L locus pJK1027
    pJK-caCas9-NatMX-Neut5L-Ade2 driveAde2 gene drive (Other)Nourseothricin Church A CRISPR-Cas9-based gene drive platform for genetic interaction analysis in Candida albicans. Nat Microbiol. 2018 Jan;3(1):73-82. doi: 10.1038/s41564-017-0043-0. Epub 2017 Oct 23. caCas9 integrating vector into the Neut5L locus with gene drive for targeting Ade2 locus pJK1027
    pJK-caCas9-NatMX-Neut5L-Leu2 driveLeu2 gene drive (Other) Church A CRISPR-Cas9-based gene drive platform for genetic interaction analysis in Candida albicans. Nat Microbiol. 2018 Jan;3(1):73-82. doi: 10.1038/s41564-017-0043-0. Epub 2017 Oct 23. caCas9 integrating vector into the Neut5L locus with gene drive for targeting Leu2 locus pjk1027
    pWS158Cas9 (Saccharomyces cerevisiae)URA3 Ellis Ellis Lab Yeast CRISPR Plasmids (unpublished) Cas9 gap repair vector - URA3 2 micron
    pWS171Cas9 (Saccharomyces cerevisiae)LEU2 Ellis Ellis Lab Yeast CRISPR Plasmids (unpublished) Cas9 gap repair vector - LEU2 2 micron
    pWS172Cas9 (Saccharomyces cerevisiae)HIS3 Ellis Ellis Lab Yeast CRISPR Plasmids (unpublished) Cas9 gap repair vector - HIS3 2 micron
    pWS173Cas9 (Saccharomyces cerevisiae)Kanamycin (select with G418) Ellis Ellis Lab Yeast CRISPR Plasmids (unpublished) Cas9 gap repair vector - KanR 2 micron
    pWS174Cas9 (Saccharomyces cerevisiae)Nourseothricin Ellis Ellis Lab Yeast CRISPR Plasmids (unpublished) Cas9 gap repair vector - NatR 2 micron
    pWS175Cas9 (Saccharomyces cerevisiae)Hygromycin Ellis Ellis Lab Yeast CRISPR Plasmids (unpublished) Cas9 gap repair vector - HygR 2 micron
    pADH99US_HIS1, FRT, pENO1, C. albicans Cas9, pMAL2, FLP, NAT 1 of 2Nourseothricin (NAT) Hernday An efficient, rapid, and recyclable system for CRISPR-mediated genome editing in Candida albicans mSphere Apr 2017, 2 (2) e00149-17 NAT-marked CAS9 expression construct; part 1 of 2 of C.alb HIS1-FLP CRISPR system. Use with pADH100-series gRNA expression constructs. pUC19
    pADH137C. albicans LEU2 1 of 2, pENO1, Cas9, NAT 1 of 2Nourseothricin (NAT) Hernday An efficient, rapid, and recyclable system for CRISPR-mediated genome editing in Candida albicans mSphere Apr 2017, 2 (2) e00149-17 NAT-marked CAS9 expression construct; part 1 of 2 of C.alb LEUpOUT CRISPR system. Use with pADH118-series gRNA expression constructs. pUC19
    pADH140C. maltosa LEU2 1 of 2, pENO1, Cas9, NAT 1 of 2Nourseothricin (NAT) Hernday An efficient, rapid, and recyclable system for CRISPR-mediated genome editing in Candida albicans mSphere Apr 2017, 2 (2) e00149-17 NAT-marked CAS9 expression construct; part 1 of 2 of C.mal LEUpOUT CRISPR system. Use with pADH143-series gRNA expression constructs. pUC19
    pDHt/sk-PCCas9 (Other)PpdcHygromycin Zhou Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system. Cell Discov. 2015 May 12;1:15007. doi: 10.1038/celldisc.2015.7. eCollection 2015. Expression of codon-optimized Cas9 with Ppdc promoter in Trichoderma reesei pDHt/sk
    pDHt/sk-PECas9-eGFP (Other)PpdcHygromycin Zhou Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system. Cell Discov. 2015 May 12;1:15007. doi: 10.1038/celldisc.2015.7. eCollection 2015. Expression of codon-optimized Cas9-eGFP fusion protein with Ppdc promoter in Trichoderma reesei pDHt/sk
  • Tag / Fusion Protein
    • EGFP (C terminal on insert)
    		•
    pDHt/sk-CCCas9 (Other)Pcbh1Hygromycin Zhou Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system. Cell Discov. 2015 May 12;1:15007. doi: 10.1038/celldisc.2015.7. eCollection 2015. Expression of codon-optimized Cas9 with Pcbh1 promoter in Trichoderma reesei pDHt/sk
    pDHt/sk-CECas9-eGFP (Other)Pcbh2Hygromycin Zhou Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system. Cell Discov. 2015 May 12;1:15007. doi: 10.1038/celldisc.2015.7. eCollection 2015. Expression of codon-optimized Cas9-eGFP fusion protein with Pcbh1 promoter in Trichoderma reesei pDHt/sk
  • Tag / Fusion Protein
    • EGFP (C terminal on insert)
    		•
    pYZ033Spura4 Boeke Construction of Designer Selectable Marker Deletions with CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized Design of Common Entry Vectors. G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 10.1534/g3.117.300363. Entry vector for S. pombe CRISPR-Cas9 system. The gRNA can be integrated easily through Gibson Assembly with the plasmid backbone digested by Not1 pMZ374
    pYZ146gRNA targeting Sp.leu1 (Schizosaccharomyces pombe)Spura4 Boeke Construction of Designer Selectable Marker Deletions with CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized Design of Common Entry Vectors. G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 10.1534/g3.117.300363. Cas9-gRNA leu1 plasmid for leu1 deletion in S. pombe pYZ033 leu1 SPBC1A4.02c
    pYZ164gRNA targeting Sp.his3 (Schizosaccharomyces pombe)Spura4 Boeke Construction of Designer Selectable Marker Deletions with CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized Design of Common Entry Vectors. G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 10.1534/g3.117.300363. Cas9-gRNA his3 plasmid for his3 deletion in S. pombe pYZ033 his3 SPBC11B10.02c
    pYZ173gRNA targeting Sp.lys9 (Schizosaccharomyces pombe)Spura4 Boeke Construction of Designer Selectable Marker Deletions with CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized Design of Common Entry Vectors. G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 10.1534/g3.117.300363. Cas9-gRNA lys9 plasmid for lys9 deletion in S. pombe pYZ033 SPBC3B8.03 SPBC3B8.03
    pDB4279Cas9 (Other) Du A Cloning-Free Method for CRISPR/Cas9-Mediated Genome Editing in Fission Yeast. G3 (Bethesda). 2018 Apr 27. pii: g3.118.200164. doi: 10.1534/g3.118.200164. A control plasmid containing the intact bsdMX marker pMZ374 (Addgene 59896)
    pDB4280Cas9 (Other) Du A Cloning-Free Method for CRISPR/Cas9-Mediated Genome Editing in Fission Yeast. G3 (Bethesda). 2018 Apr 27. pii: g3.118.200164. doi: 10.1534/g3.118.200164. A Cas9-encoding plasmid containing the 5' portion of the ura4 marker and the rrk1 promoter/leader. When linearized by NotI, it serves as the gapped vector in the split-ura4 system pMZ374 (Addgene 59896)
    pDB4281Cas9 (Other) Du A Cloning-Free Method for CRISPR/Cas9-Mediated Genome Editing in Fission Yeast. G3 (Bethesda). 2018 Apr 27. pii: g3.118.200164. doi: 10.1534/g3.118.200164. A Cas9-encoding plasmid containing the 5' portion of the bsdMX marker and the rrk1 promoter/leader. When linearized by NotI, it serves as the gapped vector in the split-bsdMX system pMZ374 (Addgene 59896)
    pIW447-CRISPR XYL2Codon optimized Cas9 (Synthetic), sgRNA expression cassette (Synthetic)URA3 Wheeldon CRISPR-Cas9-enabled genetic disruptions for understanding ethanol and ethyl acetate biosynthesis in Kluyveromyces marxianus. Biotechnol Biofuels. 2017 Jun 24;10:164. doi: 10.1186/s13068-017-0854-5. eCollection 2017. K. marxianus CRISPR Plasmid for XYL2 editing pJSK316-GPD
    pIW601-KmCRISPRCodon optimized Cas9 (Synthetic), sgRNA expression cassette (Synthetic)URA3 Wheeldon CRISPR-Cas9-enabled genetic disruptions for understanding ethanol and ethyl acetate biosynthesis in Kluyveromyces marxianus. Biotechnol Biofuels. 2017 Jun 24;10:164. doi: 10.1186/s13068-017-0854-5. eCollection 2017. K. marxianus CRISPR Plasmid for sgRNA cloning pJSK316-GPD
    bRA89Hygromycin Haber Rad51-mediated double-strand break repair and mismatch correction of divergent substrates. Nature. 2017 Apr 20;544(7650):377-380. doi: 10.1038/nature22046. Epub 2017 Apr 12. Cas9 driven by PGK1 promoter. gRNA can be cloned into the BplI sites. HPH marker pRS316
    bRA90LEU2 Haber Rad51-mediated double-strand break repair and mismatch correction of divergent substrates. Nature. 2017 Apr 20;544(7650):377-380. doi: 10.1038/nature22046. Epub 2017 Apr 12. Cas9 driven by PGK1 promoter. gRNA can be cloned into the BplI sites. LEU2 marker pRS316
    bRA66Hygromycin Haber Rad51-mediated double-strand break repair and mismatch correction of divergent substrates. Nature. 2017 Apr 20;544(7650):377-380. doi: 10.1038/nature22046. Epub 2017 Apr 12. GAL1 driven Cas9. gRNA can be cloned into BplI sites. pRS316
    bRA77LEU2 Haber Rad51-mediated double-strand break repair and mismatch correction of divergent substrates. Nature. 2017 Apr 20;544(7650):377-380. doi: 10.1038/nature22046. Epub 2017 Apr 12. GAL1 driven Cas9. gRNA can be cloned into BplI sites. pRS316
    pJH2970HIS3 Haber Cas9-mediated gene editing in Saccharomyces cerevisiae Protocol Exchange (2017) Cas9 driven by PGK1 promoter. gRNA can be cloned into the BplI sites. HIS3MX marker pRS316
    pJH2971Neomycin (select with G418) Haber Cas9-mediated gene editing in Saccharomyces cerevisiae Protocol Exchange (2017) Cas9 driven by PGK1 promoter. gRNA can be cloned into the BplI sites. KANMX marker pRS316
    pJH2972URA3 Haber Cas9-mediated gene editing in Saccharomyces cerevisiae Protocol Exchange (2017) Cas9 driven by PGK1 promoter. gRNA can be cloned into the BplI sites. URA3MX marker pRS316
    pUDP004ScTDH3amdS (acetamidase gene conferring growth on acetamide) Daran CRISPR-Cas9 mediated gene deletions in lager yeast Saccharomyces pastorianus. Microb Cell Fact. 2017 Dec 5;16(1):222. doi: 10.1186/s12934-017-0835-1. E. coli/S. cerevisiae shuttle vector carrying amd S marker and Spcas9D147Y P411T allowing cloning of ribozyme flanked g-RNA for Cas9 editing (HH-gRNA-HDV) not applicable
    pUDP012HH-gRNA-HDV targetting SeILV6 in S. pastorianus (Saccharomyces cerevisiae)ScTDH3, ScTDH3amdS (acetamidase gene conferring growth on acetamide) Daran CRISPR-Cas9 mediated gene deletions in lager yeast Saccharomyces pastorianus. Microb Cell Fact. 2017 Dec 5;16(1):222. doi: 10.1186/s12934-017-0835-1. E. coli/S. cerevisiae shuttle vector carrying amdS andSpcas9D147Y P411T and expressing a ribozyme flanked g-RNA for Cas9 editing targeting the gene SeILV6 and in S. pastorianus (HH-gRNASeILV6-HDV) pUDP004
    pUDP044polycistronic HH-gRNA-HDV-HH-gRNA-HDV array targetting SeATF1 and 2 in S. pastorianus (Saccharomyces cerevisiae)ScTDH3, ScTDH3amdS (acetamidase gene conferring growth on acetamide) Daran CRISPR-Cas9 mediated gene deletions in lager yeast Saccharomyces pastorianus. Microb Cell Fact. 2017 Dec 5;16(1):222. doi: 10.1186/s12934-017-0835-1. pUDP004 expressing a polycistronic g-RNA array for Cas9 editing targeting the genes SeATF1 and SeATF2 and Spcas9D147Y P411T in S. pastorianus (HH-gRNASeATF1-HDV-linker-HH-gRNASeATF2-HDV) pUDP004
    pCSN061S. pyogenes Cas9 codon optimized for expression in S. cerevisiae., KanMX marker expression cassette. (Other)Heterologous promoter from K. lactis (KLLA0F20031g), Heterologous TEF1 promoter from A. gossypii.TRP1 ; KanMX Verwaal CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae. Yeast. 2017 Sep 8. doi: 10.1002/yea.3278. Single copy yeast plasmid expressing Cas9 from Streptococcus pyogenes (SpCas9), codon optimized for expression in Saccharomyces cerevisiae. pRS414 NEWENTRY
    pCSN067Cpf1 from Lachnospiraceae bacterium ND2006 (LbCpf1) codon optimized for expression in S. cerevisiae. (Other), KanMX marker expression cassette. (Other)Heterologous promoter from K. lactis (KLLA0F20031g)., Heterologous TEF1 promoter from A. gossypii.TRP1 ; KanMX Verwaal CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae. Yeast. 2017 Sep 8. doi: 10.1002/yea.3278. Single copy yeast plasmid expressing Cpf1 from Lachnospiraceae bacterium ND2006 (LbCpf1), codon optimized for expression in Saccharomyces cerevisiae. pRS414
    pCSN068Cpf1 from Francisella novicida U112 (FnCpf1) codon optimized for expression in S. cerevisiae. (Other), KanMX marker expression cassette. (Other)Heterologous promoter from K. lactis (KLLA0F20031g)., Heterologous TEF1 promoter from A. gossypii.TRP1 ; KanMX Verwaal CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae. Yeast. 2017 Sep 8. doi: 10.1002/yea.3278. Single copy yeast plasmid expressing Cpf1 from Francisella novicida U112 (FnCpf1), codon optimized for expression in Saccharomyces cerevisiae. pRS414
    pYZ292LEU2 ; Sc Boeke Construction of Designer Selectable Marker Deletions with CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized Design of Common Entry Vectors. G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 10.1534/g3.117.300363. S. pombe gRNA entry vector with Cas9 - ScLEU2 marker pYZ292
    pYZ293Sp leu1 Boeke Construction of Designer Selectable Marker Deletions with CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized Design of Common Entry Vectors. G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 10.1534/g3.117.300363. S. pombe gRNA entry vector with Cas9 - Sp leu1 marker pYZ293
    pYZ294kanMX Boeke Construction of Designer Selectable Marker Deletions with CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized Design of Common Entry Vectors. G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 10.1534/g3.117.300363. S. pombe gRNA entry vector with Cas9 - KanMX marker pYZ294
    pYZ300Sp his3 Boeke Construction of Designer Selectable Marker Deletions with CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized Design of Common Entry Vectors. G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 10.1534/g3.117.300363. S. pombe gRNA entry vector with Cas9 - Sp his3 marker pYZ300
    pYZ301Sp lys9 Boeke Construction of Designer Selectable Marker Deletions with CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized Design of Common Entry Vectors. G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 10.1534/g3.117.300363. S. pombe gRNA entry vector with Cas9 - Sp lys9 marker pYZ301
    pUDE731Fncpf1 (Other)TEF1URA3 Daran FnCpf1, a novel and efficient genome editing tool for Saccharomyces cerevisiae. Nucleic Acids Research (2017), gkx1007 S. cerevisiae Episomal plasmid harboring Fncpf1 under the control of TDH3p pMEL10
    pUDC175Fncpf1 (Other)TEF1TRP1 Daran FnCpf1, a novel and efficient genome editing tool for Saccharomyces cerevisiae. Nucleic Acids Research (2017), gkx1007 S. cerevisae centromic plasmid harboring Fncpf1 under control of TEF1 promoter p414TEF1
    pUDP002hph (Other), Spcas9 D147Y P411T (Other)Ashbya gossypii (Eremothecium gossypii) TEF1, Arxula adeninivorans TEF1Hygromycin Daran Genome editing in Kluyveromyces and Ogataea yeasts using a broad-host-range Cas9/gRNA co-expression plasmid. FEMS Yeast Res. 2018 May 1;18(3). pii: 4847887. doi: 10.1093/femsyr/foy012. Broad-host-range Cas9/gRNA co-expression backbone plasmid (no gRNA) not applicable
    BB3cN_pGAP_23*_pLAT1_Cas9HH - FS23 linker - HDV (Other), human codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS) (Homo sapiens)NTC Gasser CRISPR/Cas9-mediated homology directed genome editing in Pichia pastoris (unpublished) hCas9 under control of LAT1 for direct cloning of HH-sgRNA-HDV PCR products for episomal expression in P. pastoris and selection on NTC Episomal ARS/CEN Backbone
    BB3cH_pGAP_23*_pLAT1_Cas9HH - FS23 linker - HDV (Other), human codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS) (Homo sapiens)Hygromycin Gasser CRISPR/Cas9-mediated homology directed genome editing in Pichia pastoris (unpublished) hCas9 under control of LAT1 for direct cloning of HH-sgRNA-HDV PCR products for episomal expression in P. pastoris and selection on Hyg Episomal ARS/CEN Backbone
    BB3cK_pGAP_23*_pLAT1_Cas9HH - FS23 linker - HDV (Other), human codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS) (Homo sapiens)Neomycin (select with G418) Gasser CRISPR/Cas9-mediated homology directed genome editing in Pichia pastoris (unpublished) hCas9 under control of LAT1 for direct cloning of HH-sgRNA-HDV PCR products for episomal expression in P. pastoris and selection on G418 Episomal ARS/CEN Backbone
    BB3cK_pGAP_23*_pTEF_Cas9HH - FS23 linker - HDV (Other), human codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS) (Homo sapiens)Neomycin (select with G418) Gasser CRISPR/Cas9-mediated homology directed genome editing in Pichia pastoris (unpublished) hCas9 under control of Tef1 for direct cloning of HH-sgRNA-HDV PCR products and episomal expression in P. pastoris and G418 selection Episomal ARS/CEN Backbone
    BB3cK_pGAP_23*_pPFK300_Cas9HH - FS23 linker - HDV (Other), human codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS) (Homo sapiens)Neomycin (select with G418) Gasser CRISPR/Cas9-mediated homology directed genome editing in Pichia pastoris (unpublished) hCas9 under control of pPFK300 for direct cloning of HH-sgRNA-HDV PCR products for episomal expression in P. pastoris and selection on G418 Episomal ARS/CEN Backbone
    BB3cN_pGAP_23*_pPFK300_Cas9HH - FS23 linker - HDV (Other), human codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS) (Homo sapiens)NTC Gasser CRISPR/Cas9-mediated homology directed genome editing in Pichia pastoris (unpublished) hCas9 under control of pPFK300 for direct cloning of HH-sgRNA-HDV PCR products for episomal expression in P. pastoris and selection on NTC Episomal ARS/CEN Backbone
    BB3cH_pGAP_23*_pPFK300_Cas9HH - FS23 linker - HDV (Other), human codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS) (Homo sapiens)Hygromycin Gasser CRISPR/Cas9-mediated homology directed genome editing in Pichia pastoris (unpublished) hCas9 under control of pPFK300 for direct cloning of HH-sgRNA-HDV PCR products for episomal expression in P. pastoris and selection on Hyg Episomal ARS/CEN Backbone
    pCfB4906Cas9 (Other)Hygromycin Borodina EasyCloneYALI: CRISPR/Cas9-Based Synthetic Toolbox for Engineering of the Yeast Yarrowia lipolytica. Biotechnol J. 2018 Jan 29. doi: 10.1002/biot.201700543. EasyCloneYALI system-based yeast integrative vector expressing Cas9 carrying hygromycin resistance marker, integration into Yarrowia lipolytica chromosomal location IntB, amp resistance Unknown
    pCfB6364Cas9 (Other)D-serine marker; enables growth on D-serine Borodina EasyCloneYALI: CRISPR/Cas9-Based Synthetic Toolbox for Engineering of the Yeast Yarrowia lipolytica. Biotechnol J. 2018 Jan 29. doi: 10.1002/biot.201700543. EasyCloneYALI system-based yeast integrative vector expressing Cas9 carrying D-serine marker, integration into Yarrowia lipolytica ku70 locus, amp resistance Unknown
    pHCas9-NoursCas9 (Other)TEF1URA3 Yang Unraveling the genetic basis of fast l-arabinose consumption on top of recombinant xylose-fermenting Saccharomyces cerevisiae. Biotechnol Bioeng. 2018 Sep 10. doi: 10.1002/bit.26827. Escherichia coli- S. cerevisiae shuttle plasmid harbors a Cas9 gene, a natMX6 and URA3 selection markers used in S. cerevisiae pSH47
    pV1025CaCas9 (Synthetic)Ampicillin Fink A CRISPR system permits genetic engineering of essential genes and gene families. Sci Adv. 2015;1(3):e1500248. Expression of Candida albicans codon-optimized Cas9 (CaCas9) - integrates at ENO1 (Part of Duet system) pV941
    pV1093CaCas9/sgRNA-BsmBI stuffer (Synthetic)clonNAT Fink A CRISPR system permits genetic engineering of essential genes and gene families. Sci Adv. 2015;1(3):e1500248. CaCas9/gRNA Solo entry plasmid for cloning guides - contains stuffer with BsmBI sites - integrates at ENO1 pV1088/pV1090
    pV1200CaCas9/sgRNA-BsmBI stuffer (Synthetic)Ampicillin Fink A CRISPR system permits genetic engineering of essential genes and gene families. Sci Adv. 2015;1(3):e1500248. CaCas9/gRNA Solo entry plasmid for cloning guides - contains stuffer with BsmBI sites - integrates at ENO1 - Recyclable for serial mutagenesis (Sap2-FLP) pV1025/pV1093
    pV1393CaCas9/sgRNA-BsmBI stuffer (Synthetic)Ampicillin Fink New CRISPR Mutagenesis Strategies Reveal Variation in Repair Mechanisms among Fungi. mSphere. 2018 Apr 25;3(2). pii: 3/2/e00154-18. doi: 10.1128/mSphere.00154-18. Print 2018 Apr 25. CaCas9/gRNA Solo entry plasmid for cloning guides - contains stuffer with BsmBI sites - inserts at Neut5L - Recyclable for serial mutagenesis (Sap2-FLP) pV1390
    pV1524CaCas9/sgRNA-BsmBI stuffer (Synthetic)Ampicillin Fink New CRISPR Mutagenesis Strategies Reveal Variation in Repair Mechanisms among Fungi. mSphere. 2018 Apr 25;3(2). pii: 3/2/e00154-18. doi: 10.1128/mSphere.00154-18. Print 2018 Apr 25. CaCas9/gRNA Solo entry plasmid for cloning guides - contains stuffer with BsmBI sites - inserts at Neut5L - Recyclable for serial mutagenesis (Mal2-FLP) pV1393
    pV1539CaCas9/sgRNA-BsmBI stuffer (Synthetic)Ampicillin Fink New CRISPR Mutagenesis Strategies Reveal Variation in Repair Mechanisms among Fungi. mSphere. 2018 Apr 25;3(2). pii: 3/2/e00154-18. doi: 10.1128/mSphere.00154-18. Print 2018 Apr 25. Genedrive entry vector for Candida albicans pV1524
    pV1326CaCas9/sgRNA-BsmBI stuffer (Synthetic)URA3 ; clonNAT Fink New CRISPR Mutagenesis Strategies Reveal Variation in Repair Mechanisms among Fungi. mSphere. 2018 Apr 25;3(2). pii: 3/2/e00154-18. doi: 10.1128/mSphere.00154-18. Print 2018 Apr 25. S. cerevisiae and C. glabrata Solo CRISPR vector, marked with URA3 and NatR pRS416
    pV1382CaCas9/sgRNA-BsmBI stuffer (Synthetic)URA3 ; clonNAT Fink New CRISPR Mutagenesis Strategies Reveal Variation in Repair Mechanisms among Fungi. mSphere. 2018 Apr 25;3(2). pii: 3/2/e00154-18. doi: 10.1128/mSphere.00154-18. Print 2018 Apr 25. S. cerevisiae and C. glabrata Solo CRISPR vector, marked with URA3 and NatR pRS416
    pV1464CaCas9/sgRNA-BsmBI stuffer (Synthetic)URA3 ; clonNAT Fink New CRISPR Mutagenesis Strategies Reveal Variation in Repair Mechanisms among Fungi. mSphere. 2018 Apr 25;3(2). pii: 3/2/e00154-18. doi: 10.1128/mSphere.00154-18. Print 2018 Apr 25. N. castellii Solo CRISPR vector, marked with URA3 and NatR pRS416
    pZS157 CRISPEY RT/Cas9SpCas9, Ec86-RT (Other)GAL1-GAL10HIS3 Fraser Functional Genetic Variants Revealed by Massively Parallel Precise Genome Editing. Cell. 2018 Sep 18. pii: S0092-8674(18)31118-8. doi: 10.1016/j.cell.2018.08.057. Yeast Integration Plasmid for galactose inducible Ec86-RT and SpCas9 expression pRS403
    eGFP L202 gRNAeGFP L202 gRNA (Other)U6 Harris A panel of eGFP reporters for single base editing by APOBEC-Cas9 editosome complexes. Sci Rep. 2019 Jan 24;9(1):497. doi: 10.1038/s41598-018-36739-9. gRNA that guides Cas9 and APOBEC complexes to L202 in eGFP. MLM3636
    eGFP L138 gRNAeGFP L138 gRNA (Other)U6 Harris A panel of eGFP reporters for single base editing by APOBEC-Cas9 editosome complexes. Sci Rep. 2019 Jan 24;9(1):497. doi: 10.1038/s41598-018-36739-9. gRNA that guides Cas9 and APOBEC complexes to L138 in eGFP. MLM3636
    pAH235humanized Streptococcus pyogenes Cas9 (Synthetic)nmt41S.pombe ura4 Tanaka Short-Homology-Mediated CRISPR/Cas9-Based Method for Genome Editing in Fission Yeast. G3 (Bethesda). 2019 Feb 12. pii: g3.118.200976. doi: 10.1534/g3.118.200976. Cas9 expression controlled by the nmt41 promoter pSLF273
    pAH237gRNA cassette (Schizosaccharomyces pombe), humanized Streptococcus pyogenes Cas9 (Schizosaccharomyces pombe)S.pombe rrk1, nmt41LEU2 Tanaka Short-Homology-Mediated CRISPR/Cas9-Based Method for Genome Editing in Fission Yeast. G3 (Bethesda). 2019 Feb 12. pii: g3.118.200976. doi: 10.1534/g3.118.200976. Expression of both nmt41p-cas9 and rrk1p-gRNA (LEU2 marker) for CRISPR genome editing in fission yeast pAH233
    pAH243humanized Streptococcus pyogenes Cas9 (Schizosaccharomyces pombe), gRNA expression module (Schizosaccharomyces pombe)nmt41, S.pombe rrk1S.pombe ura4 Tanaka Short-Homology-Mediated CRISPR/Cas9-Based Method for Genome Editing in Fission Yeast. G3 (Bethesda). 2019 Feb 12. pii: g3.118.200976. doi: 10.1534/g3.118.200976. Expression of both nmt41p-cas9 and rrk1p-gRNA (ura4 marker) for CRISPR genome editing in fission yeast pSLF273
    pUCC001HH-BsaI (Synthetic)TDH3 promoter from S.cerevisiae (already present in backbone)Hygromycin Morrissey Biological Parts for Kluyveromyces marxianus Synthetic Biology Front. Bioeng. Biotechnol., 07 May 2019 Multi-species CRISPR/Cas9 coexpression system, optimized for Golden Gate cloning of gRNA targets pUDP002
    NM9-SpCas9-NLS3SpCas9-NLS3 (Other)pPGK1Neomycin (select with G418) Zhao Development of a CRISPR/Cas9 system for high efficiency multiplexed gene deletion in Rhodosporidium toruloides. Biotechnol Bioeng. 2019 Apr 30. doi: 10.1002/bit.27001. Expresses R. toruloides codon-optimized SpCas9 under PGK1 promoter pCAMBRIA
    pSDMA65Hygromycin (HYG) (Synthetic)ACT1Hygromycin Fraser Targeted Genome Editing via CRISPR in the Pathogen Cryptococcus neoformans. (unpublished) Codon optimised Streptococcus pyogenes CAS9 ORF regulated by the Cryptococcus neoformans TEF1 promoter and terminator. pBlueScript II SK(-)
    JME4390LEU2 Rossignol A set of Yarrowia lipolytica CRISPR/Cas9 vectors for exploiting wild-type strain diversity. Biotechnol Lett. 2020 Jan 23. pii: 10.1007/s10529-020-02805-4. doi: 10.1007/s10529-020-02805-4. LEU2ex_CrisprCas9-yl_RFP: CAS9 vector with LEU2ex marker for gRNA cloning using GoldenGate pSB1A3
    JME4393LYS5 Rossignol A set of Yarrowia lipolytica CRISPR/Cas9 vectors for exploiting wild-type strain diversity. Biotechnol Lett. 2020 Jan 23. pii: 10.1007/s10529-020-02805-4. doi: 10.1007/s10529-020-02805-4. LYS5ex_CrisprCas9-yl_RFP: CAS9 vector with LYS5ex marker for gRNA cloning using GoldenGate pSB1A3
    JME4472URA3 Rossignol A set of Yarrowia lipolytica CRISPR/Cas9 vectors for exploiting wild-type strain diversity. Biotechnol Lett. 2020 Jan 23. pii: 10.1007/s10529-020-02805-4. doi: 10.1007/s10529-020-02805-4. URA3ex_CrisprCas9-yl_RFP: CAS9 vector with URA3ex marker for gRNA cloning using GoldenGate pSB1A3
    JME4580Hygromycin Rossignol A set of Yarrowia lipolytica CRISPR/Cas9 vectors for exploiting wild-type strain diversity. Biotechnol Lett. 2020 Jan 23. pii: 10.1007/s10529-020-02805-4. doi: 10.1007/s10529-020-02805-4. HPHex_CrisprCas9-yl_RFP: CAS9 vector with Hygromycinex marker for gRNA cloning using GoldenGate pSB1A3
    JME4599Nourseothricin Rossignol A set of Yarrowia lipolytica CRISPR/Cas9 vectors for exploiting wild-type strain diversity. Biotechnol Lett. 2020 Jan 23. pii: 10.1007/s10529-020-02805-4. doi: 10.1007/s10529-020-02805-4. NATex_CrisprCas9-yl_RFP: CAS9 vector with Nourseothricin ex marker for gRNA cloning using GoldenGate pSB1A3
    pCP-tRNAcassette for the expression of the sgRNA from the C. parapsilosis RNA pol II GAPDH promoter (Synthetic)SAT1 (resistance to Nourseothricin) Butler Plasmid-Based CRISPR-Cas9 Gene Editing in Multiple Candida Species. mSphere. 2019 Mar 13;4(2). pii: 4/2/e00125-19. doi: 10.1128/mSphere.00125-19. CRISPR-Cas9 system for genetic manipulation of Candida parapsilosis, C. orthopsilosis, and C. metapsilosis pUC57
    pCT-tRNAcassette for the expression of the sgRNA from the Ashbya gossypii RNA pol II TEF1 promoter (Synthetic)SAT1 (resistance to Nourseothricin) Butler Plasmid-Based CRISPR-Cas9 Gene Editing in Multiple Candida Species. mSphere. 2019 Mar 13;4(2). pii: 4/2/e00125-19. doi: 10.1128/mSphere.00125-19. CRISPR-Cas9 system for genetic manipulation of Candida tropicalis pAYCU268
    pAEF5Hygromycin Fischer Yeast Protocols 4th Edition--A Versatile Protocol to Generate Translocations in Yeast Genomes using CRISPR/Cas9 (unpublished) Plasmid encoding the Cas9 gene and a gRNA expression cassette allowing to clone a single or multiple gRNAs for DSBs induction in yeast. Constructed by Aubin Fleiss pGangZhao110

    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 Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    pRS315e_pGal-dCas9-SH3-pGal-PmCDA1-SHLSpCas9 (Other), PmCDA1 (Other)pGal1, pGal10LEU2 Kondo Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems. Science. 2016 Aug 4. pii: aaf8729. Expresses dCas9-SH3 and PmCDA1-SHL in yeast cells pRS315
    pRS315e_pGal-dCas9-PmCDA1SpCas9 (Other)pGal1LEU2 Kondo Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems. Science. 2016 Aug 4. pii: aaf8729. Expresses dCas9-PmCDA1 in yeast cells pRS315
    pRS315e_pGal-nCas9(D10A)-PmCDA1SpCas9 (Other)pGal1LEU2 Kondo Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems. Science. 2016 Aug 4. pii: aaf8729. Expresses nCas9(D10A)-PmCDA1 in yeast cells pRS315
    pJT112_GalL_nCDA1-xBE3nCDA1-xBE3 (Other)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor nCDA1-xBE3 in yeast cells p415
    pJT25_GalL_BE3BE3 (Other)GalLLEU2 Bock Engineering of high-precision base editors for site-specific single nucleotide replacement. Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8. Expresses BE3 in yeast cells p415
    pJT78_GalL_nCDA1Δ195-BE3nCDA1Δ195-BE3 (Other)GalLLEU2 Bock Engineering of high-precision base editors for site-specific single nucleotide replacement. Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8. Expresses nCDA1Δ195-BE3 in yeast cells p415
    pJT79_GalL_nCDA1Δ192-BE3nCDA1Δ192-BE3 (Other)GalLLEU2 Bock Engineering of high-precision base editors for site-specific single nucleotide replacement. Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8. Expresses nCDA1Δ192-BE3 in yeast cells p415
    pJT82_GalL_nCDA1Δ198-BE3nCDA1Δ198-BE3 (Other)GalLLEU2 Bock Engineering of high-precision base editors for site-specific single nucleotide replacement. Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8. Expresses nCDA1Δ198-BE3 in yeast cells p415
    pJT84_GalL_nCDA1Δ194-BE3nCDA1Δ194-BE3 (Other)GalLLEU2 Bock Engineering of high-precision base editors for site-specific single nucleotide replacement. Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8. Expresses nCDA1Δ194-BE3 in yeast cells p415
    pJT85_GalL_nCDA1Δ193-BE3nCDA1Δ193-BE3 (Other)GalLLEU2 Bock Engineering of high-precision base editors for site-specific single nucleotide replacement. Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8. Expresses nCDA1Δ193-BE3 in yeast cells p415
    pJT90_GalL_nCDA1Δ190-BE3nCDA1Δ190-BE3 (Other)GalLLEU2 Bock Engineering of high-precision base editors for site-specific single nucleotide replacement. Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8. Expresses nCDA1Δ190-BE3 in yeast cells p415
    pJT92_GalL_nCDA1Δ188-BE3nCDA1Δ188-BE3 (Other)GalLLEU2 Bock Engineering of high-precision base editors for site-specific single nucleotide replacement. Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8. Expresses nCDA1Δ188-BE3 in yeast cells p415
    pJT113_GalL_cCDA1-xBE3cCDA1-xBE3 (Other)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor cCDA1-xBE3 in yeast cells p415
    pJT121_GalL_nCDA1-VQRBE3nCDA1-VQRBE3 (Other)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor nCDA1-VQRBE3 in yeast cells p415
    pJT122_GalL_cCDA1-VQRBE3cCDA1-VQRBE3 (Other)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor cCDA1-VQRBE3 in yeast cells p415
    pJT130_GalL_nCDA1-NGBE3nCDA1-NGBE3 (Other)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor nCDA1-NGBE3 in yeast cells p415
    pJT131_GalL_cCDA1-NGBE3cCDA1-NGBE3 (Other)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor cCDA1-NGBE3 in yeast cells p415
    pJT139_GalL_nCDA1-VRERBE3nCDA1-VRERBE3 (Other)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor nCDA1-VRERBE3 in yeast cells p415
    pJT166_GalL_A3A-BE3A3A-BE3 (Homo sapiens)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor A3A-BE3 in yeast cells p415
    pJT169_GalL_A3AΔ186-BE3A3AΔ186-BE3 (Homo sapiens)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor A3AΔ186-BE3 in yeast cells p415
    pJT170_GalL_A3AΔ182-BE3A3AΔ182-BE3 (Homo sapiens)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor A3AΔ182-BE3 in yeast cells p415
    pJT171_GalL_A3AΔ178-BE3A3AΔ178-BE3 (Homo sapiens)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor A3AΔ178-BE3 in yeast cells p415
    pJT173_GalL_A3A-R128A-BE3A3A(R128A)-BE3 (Homo sapiens)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor A3A-R128A-BE3 in yeast cells p415
    pJT174_GalL_A3A-Y130F-BE3A3A(Y130F)-BE3 (Homo sapiens)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor A3A-Y130F-BE3 in yeast cells p415
    pJT175_GalL_A3A(Y130F)Δ186-BE3A3A(Y130F)Δ186-BE3 (Homo sapiens)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor A3A(Y130F)Δ186-BE3 in yeast cells p415
    pJT176_GalL_eA3A-BE3A3A(N57G)-BE3 (Homo sapiens)GalLLEU2 Bock Expanding the genome-targeting scope and the site selectivity of high-precision base editors. Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. Expressing base editor eA3A-BE3 in yeast cells p415

    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).

    ID Plasmid Purpose Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    79616pRS415_pGal-nCas9(H840A)Expresses nCas9(H840A) in yeast cellsSpCas9 (Other)pGal1LEU2 Kondo Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems. Science. 2016 Aug 4. pii: aaf8729. Expresses nCas9(H840A) in yeast cells pRS415
    79617pRS315e_pGal-nCas9(D10A)-PmCDA1Expresses nCas9(D10A)-PmCDA1 in yeast cellsSpCas9 (Other)pGal1LEU2 Kondo Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems. Science. 2016 Aug 4. pii: aaf8729. Expresses nCas9(D10A)-PmCDA1 in yeast cells pRS315
    79618pRS415_pGal-nCas9(D10A)Expresses nCas9(D10A) in yeast cellsSpCas9 (Other)pGal1LEU2 Kondo Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems. Science. 2016 Aug 4. pii: aaf8729. Expresses nCas9(D10A) in yeast cells pRS415

    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 Selectable Marker PI Publication Hidden Extra Search Info
    pTPGI_dCas9_VP64dCas9_VP64 (codon-optimized for expression in S. cerevisiae) (Saccharomyces cerevisiae)pTPGI (galactose+aTc inducible)TRP1 Lu Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas. ACS Synth Biol. 2013 Sep 11. encodes yeast-optimized dCas9_VP64 synthetic transcription factor pTPGI (pRS304)
    pJZC519dCas9-VP64 (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-VP64 expression plasmid pNH605
    pJZC620MCP-VP64, PCP-VP64, dCas9pAdh, pAdh, 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. Expresses dCas9, MCP-VP64, and PCP-VP64 in Yeast cells pNH605
    pJZC638MCP-VP64, dCas9 (Other)pAdh, pGal10LEU2 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. Expresses MCP-VP64 and dCas9 in Yeast cells pNH605
    pJZC548sgRNA + 1x PP7SNR52URA3 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 PP7 for yeast cells pRS416
    pJZC595MCP-VP64, dCas9pAdh, Tdh3LEU2 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. Expresses MCP-VP64 and dCas9 in Yeast cells pNH605
    pAG414GPD-dCas9-VPRdCas9-VPR (Saccharomyces cerevisiae)GPDTRP1 Church Highly efficient Cas9-mediated transcriptional programming. Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312. pAG414 series plasmid with GPD promoter driving expression of dCas9-VPR; cerevisiae vector pAG414GPD
    pRS416-Gal4-dCas9-VP64Gal4-dCas9-VP64 (Saccharomyces cerevisiae)Tef1URA3 Davis Dissecting the Genetic Basis of a Complex cis-Regulatory Adaptation. PLoS Genet. 2015 Dec 29;11(12):e1005751. doi: 10.1371/journal.pgen.1005751. eCollection 2015 Dec. This plasmid contains a Cas9 Activator for yeast. The activator is about 1.2-1.8 X more potent than just dCas9-VP64 fusion in yeast. It is on a single copy CEN/ARS plasmid with Ura marker, pRS416. pRS416
    dCas9v2dCas9-VPR (Saccharomyces cerevisiae)URA3 Nielsen Multiplexed CRISPR/Cas9 Genome Editing and Gene Regulation using Csy4 in Saccharomyces cerevisiae. ACS Synth Biol. 2017 Nov 21. doi: 10.1021/acssynbio.7b00259. TetR inducible dCas9-VPR plasmid with pRPR-NotI-tRPR1 Csy4-ready site pDTU-113
    pCRISPRa_VPR_ylCodon optimized dCas9-VPR (Synthetic), sgRNA expression cassette (Synthetic)UAS1B8-TEF(136), SCR1'-tRNALEU2 Wheeldon Multiplexed CRISPR activation of cryptic sugar metabolism enables Yarrowia lipolytica growth on cellobiose. Biotechnol J. 2018 May 5:e1700584. doi: 10.1002/biot.201700584. CRISPR-dCas9-VPR vector for Yarrowia lipolytica, expressing dCas9-VPR and AvrII site for sgRNA insertion pUC19

    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 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
    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
    pCRISPRi_Mxi1_ylCodon optimized dCas9-Mxi1 (Synthetic), sgRNA expression cassette (Synthetic)UAS1B8-TEF(136), SCR1'-tRNALEU2 Wheeldon CRISPRi repression of nonhomologous end-joining for enhanced genome engineering via homologous recombination in Yarrowia lipolytica. Biotechnol Bioeng. 2017 Aug 19. doi: 10.1002/bit.26404. CRISPR-dCas9-Mxi1 vector for Yarrowia lipolytica, expressing dCas9-Mxi1 and AvrII site for sgRNA insertion pUC19
    pCRISPRi_Mxi1_yl_NHEJCodon optimized dCas9-Mxi1 (Synthetic), KU70 sgRNA expression cassette (Synthetic), KU80a sgRNA expression cassette (Synthetic), KU80b sgRNA expression cassette (Synthetic)UAS1B8-TEF(136), SCR1'-tRNA, SCR1'-tRNA, SCR1'-tRNALEU2 Wheeldon CRISPRi repression of nonhomologous end-joining for enhanced genome engineering via homologous recombination in Yarrowia lipolytica. Biotechnol Bioeng. 2017 Aug 19. doi: 10.1002/bit.26404. CRISPRi vector for Yarrowia lipolytica, repressing KU70 and KU80 for enhanced HR pUC19
    pRS143NAT Shapiro A CRISPR Interference Platform for Efficient Genetic Repression in Candida albicans. mSphere. 2019 Feb 13;4(1). pii: 4/1/e00002-19. doi: 10.1128/mSphere.00002-19. CRISPRi plasmid for use in Candida albicans. Contains dCas9 and NEUT5L integration site and the sgRNA cloning site (SNR52 promoter, PacI sites, sgRNA tail) pJK-caCas9-NatMX-Neut5L
    pRS159NAT Shapiro A CRISPR Interference Platform for Efficient Genetic Repression in Candida albicans. mSphere. 2019 Feb 13;4(1). pii: 4/1/e00002-19. doi: 10.1128/mSphere.00002-19. CRISPRi plasmid for use in Candida albicans. Contains dCas9-Mxi1 and NEUT5L integration site and the sgRNA cloning site (SNR52 promoter, PacI sites, sgRNA tail) pRS143
    pMM585dCas9 (Saccharomyces cerevisiae) McClean Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. Part Plasmid for dCas9, Part 3a(coding sequence) pYTK001
    pMM586dCas9 (Saccharomyces cerevisiae) McClean Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. Part Plasmid for dCas9, Part 3(coding sequence) pYTK001
    pMM587dCas9 (Saccharomyces cerevisiae) McClean Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. Part Plasmid for dCas9, Part 3b(coding sequence) pYTK001
    pMM729Mxi1-dCas9 (Saccharomyces cerevisiae) McClean Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. Part Plasmid for Mxi1-dCas9, Part 3(coding sequence) pYTK001
    pMM765dCas9 NLS Stop (Saccharomyces cerevisiae) McClean Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. Part Plasmid for dCas9 NLS Stop, Part 3(coding sequence) pYTK001
    pMM766dCas9 NLS Stop (Saccharomyces cerevisiae) McClean Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. Part Plasmid for dCas9 NLS Stop, Part 3b(coding sequence) pYTK001
    pMM767dCas9 NLS (Saccharomyces cerevisiae) McClean Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. Part Plasmid for dCas9 NLS, Part 3(coding sequence) pYTK001
    pMM768dCas9 NLS (Saccharomyces cerevisiae) McClean Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. Part Plasmid for dCas9 NLS, Part 3b(coding sequence) pYTK001
    pMM769dCas9 NLS (Saccharomyces cerevisiae) McClean Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. Part Plasmid for dCas9 NLS, Part 3a(coding sequence) pYTK001
    p415:iCas9mTN3-GGS6-dCas9 (Synthetic)GalLLEU2 Wang RNA-Guided Recombinase-Cas9 Fusion Targets Genomic DNA Deletion and Integration. CRISPR J. 2019 Aug;2:209-222. doi: 10.1089/crispr.2019.0013. Expression of mTN3-GGS6-dCas9 (iCas9) in yeast. p415

    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 Selectable Marker PI Publication Hidden Extra Search Info
    3xFLAG-dCas9/pTEF1p-CYC1t3xFLAG-dCas9 (Synthetic)TEF1 promoterTRP1 Fujii An enChIP system for the analysis of bacterial genome functions. BMC Res Notes. 2018 Jun 14;11(1):387. doi: 10.1186/s13104-018-3486-3. Expresses 3xFLAG-dCas9 in budding yeast for enChIP analysis to purify specific genomic regions of interest. p414

    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)
    		pRPR1_gRNA_
    handle_RPR1t    		 pRPR1 HindIII S. cerevisiae LEU2 none, need Cas9 plasmid Lu

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