CRISPR/Cas Plasmids: Activate Gene Expression

A catalytically inactive Cas9 (dCas9) fused to a transcription activator peptide can increase transcription of a gene. Design your gRNA sequence to direct the dCas9-activator to a specific genomic sequence. Potential target locations can include promoter regions and regulatory 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-activator.

Mammalian

Plasmid	Gene/Insert	Promoter	Selectable Marker	PI	Publication	Hidden Extra Search Info
pMSCV-LTR-dCas9-VP64-BFP	dCas9-VP64-BFP fusion (Homo sapiens), Puromycin resistance	LTR, PGK	Puromycin	Qi	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 MSCV LTR promoter, dCas9 that is fused to 2x NLS, VP64 and tagBFP MSCV-puro
pMSCV-LTR-dCas9-p65AD-BFP	dCas9-p65AD-BFP fusion (Homo sapiens), Puromycin resistance	LTR, PGK	Puromycin	Qi	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 MSCV LTR promoter, dCas9 that is fused to 2x NLS, p65 activation domain and tagBFP MSCV-puro
pcDNA-dCas9-VP64	dCas9-VP64 (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) fused to VP64 transactivator domain in mammalian cells pcDNA3.1
Cas9m2-VP64	Cas9m2-VP64 (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.	Cas9m2 Activator pcDNA3.3_TOPO
Cas9m3-VP64	Cas9m3-VP64 (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.	Cas9m3 Activator pcDNA3.3_TOPO
Cas9m4-VP64	Cas9m4-VP64 (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.	Cas9m4 Activator pcDNA3.3_TOPO
pSL690	dCas9-VP64 (Synthetic)	CMV		Joung	CRISPR RNA-guided activation of endogenous human genes. Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2598.	Expresses dCas9-VP64 fusion unknown
pMLM3705	codon optimized dCas9-VP64 (Synthetic)	CMV		Joung	CRISPR RNA-guided activation of endogenous human genes. Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2598.	Expresses mammalian cell codon-optimized dCas9-VP64 pJDS246
pAC1-pCR8-dCas9VP48	dCas9(D10A;H840A) fusion with VP48 activation domain (Synthetic)			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.	dCas9VP48 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
pAC147-pCR8-dCas9VP64	dCas9(D10A;H840A) fusion with VP64 activation domain (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.	dCas9VP64 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
pAC148-pCR8-dCas9VP96	dCas9(D10A;H840A) fusion with VP96 activation domain (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.	dCas9VP96 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
pAC149-pCR8-dCas9VP160	dCas9(D10A;H840A) fusion with VP160 activation domain (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.	dCas9VP160 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
pAC91-pmax-dCas9VP64	dCas9(D10A;H840A) fusion with VP64 activation domain (Homo sapiens)	CAGGS		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.	dCas9VP64 on pmax expression vector pmax-DEST (Addgene: 48222)
pAC92-pmax-dCas9VP96	dCas9(D10A;H840A) fusion with VP96 activation domain (Homo sapiens)	CAGGS		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.	dCas9VP96 on pmax expression vector pmax-DEST (Addgene: 48222)
pAC93-pmax-dCas9VP160	dCas9(D10A;H840A) fusion with VP160 activation domain (Homo sapiens)	CAGGS		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.	dCas9VP160 on pmax expression vector pmax-DEST (Addgene: 48222)
pAC94-pmax-dCas9VP160-2A-puro	dCas9(D10A;H840A) fusion with VP160 activation domain followed by 2A-puro (Homo sapiens)	CAGGS	Puromycin	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.	dCas9VP160-2A-puro (puro-selectable) on pmax expression vecor. Note: This is being tested. pmax-DEST (Addgene: 48222)
pAC95-pmax-dCas9VP160-2A-neo	dCas9(D10A;H840A) fusion with VP160 activation domain followed by 2A-neo (Homo sapiens)	CAGGS	Neomycin (select with G418)	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.	dCas9VP160-2A-neo (neo/G418-selectable) on pmax expression vector. Note: This is being tested. pmax-DEST (Addgene: 48222)
pAC2-dual-dCas9VP48-sgExpression	dCas9VP48 (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.	Dual expression construct expressing both dCas9VP48 and sgRNA from separate promoters pX335 (Addgene #42335)
pAC5-dual-dCas9VP48-sgTetO	dCas9VP48 and sgTetO (Synthetic)			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.	Dual expression construct expressing both dCas9VP48 and sgTetO from separate promoters pAC2-dual-dCas9VP48-sgExpression (Addgene #48236)
pAC152-dual-dCas9VP64-sgExpression	dCas9 (Synthetic)			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.	Dual expression construct expressing both dCas9VP64 and sgRNA from separate promoters pX335 (Addgene #42335) Tags / Fusion Proteins HA-Tag (N terminal on insert) VP64 (C terminal on insert)
pAC153-dual-dCas9VP96-sgExpression	dCas9 (Synthetic)			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.	Dual expression construct expressing both dCas9VP96 and sgRNA from separate promoters pX335 (Addgene #42335) Tags / Fusion Proteins HA-Tag (N terminal on insert) VP96 (C terminal on insert)
pAC154-dual-dCas9VP160-sgExpression	dCas9 (Synthetic)			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.	Dual expression construct expressing both dCas9VP160 and sgRNA from separate promoters pX335 (Addgene #42335) Tags / Fusion Proteins HA-Tag (N terminal on insert) VP160 (C terminal on insert)
M-SPn-VP64	Cas9-VP64, nuclease-null (Other)	CMV	Neomycin (select with G418)	Church	Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.	Mammalian SP-VP64 nuclease-null Cas9 activator expression, human optimized pcDNA3.3 TOPO
M-ST1n-VP64	Cas9-VP64, nuclease-null (Other)	CMV	Neomycin (select with G418)	Church	Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.	Mammalian ST1-VP64 nuclease-null Cas9 activator expression, human optimized pcDNA3.3 TOPO
M-NMn-VP64	Cas9-VP64, nuclease-null (Other)	CMV	Neomycin (select with G418)	Church	Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.	Mammalian NM-VP64 nuclease-null Cas9 activator expression, human optimized pcDNA3.3 TOPO
pCMV_dCas9_VP64	dCas9_VP64 (human-codon-optimized) (Homo sapiens)	CMV		Lu	Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas. ACS Synth Biol. 2013 Sep 11.	encodes human-optimized dCas9_VP64 synthetic transcription factor phi-Yellow-Dest
pHAGE TRE dCas9-VP64	dCas9 (Other)	TRE	Neomycin (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-VP64 lentiviral expression vector pHAGE
pHAGE EF1α dCas9-VP64	dCas9 (Other)	EF1alpha	Puromycin	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-VP64 lentiviral expression vector pHAGE
pLV hUbC-dCas9 VP64-T2A-GFP	humanized dead Cas9 VP64 T2A GFP (Other)	hUbC		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-VP64 and GFP FUGW
CMVp-dCas9-3xNLS-VP64 (Construct 1)	dCas9 (Homo sapiens)	CMV/hUBC		Lu	Multiplexed and Programmable Regulation of Gene Networks with an Integrated RNA and CRISPR/Cas Toolkit in Human Cells. Mol Cell. 2014 May 14. pii: S1097-2765(14)00355-4. doi: 10.1016/j.molcel.2014.04.022.	Expresses taCas9 in Mammalian cells for transactivating endogenous and synthetic promoters. The backbone is a lentiviral vector. pFUGw (Addgene id: 25870)
pLV hUbC-VP64 dCas9 VP64-T2A-GFP	humanized VP64 dead Cas9 VP64 T2A GFP (Other)			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 fused to two copies of VP64 and GFP FUGW
pcDNA3.1-CibN-dCas9	CibN-dCas9 (Arabidopsis thaliana)	CMV	Neomycin (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-CibN	dCas9-CibN (Arabidopsis thaliana)	CMV	Neomycin (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-CibN	dCas9-CibN (Arabidopsis thaliana)	CMV	Neomycin (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
pHRdSV40-dCas9-10xGCN4_v4-P2A-BFP	Cas9 dead			Vale	A Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging. Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. doi: 10.1016/j.cell.2014.09.039.	Expressed a nuclease dead Cas9 tagged with 10 copies of the GCN4 peptide v4 and BFP. This plasmid is part of the SunTag system for gene activation pHR
pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS	scFv-GCN4			Vale	A Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging. Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. doi: 10.1016/j.cell.2014.09.039.	The plasmid encodes a antibody that binds to the GCN4 peptide from the SunTag system, and is fused to a transcriptional activation domain VP64 pHR
pHRdSV40-NLS-dCas9-24xGCN4_v4-NLS-P2A-BFP-dWPRE	dCas9	dSV40 Promoter		Vale	A Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging. Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. doi: 10.1016/j.cell.2014.09.039.	dCas9 fused to 24 copies of the GCN4 peptide v4, which is part of the SunTag system pHR
pcDNA-dCas9-FLp300	S.pyogenes dCas9 with c-terminal full length human p300 (aa 2-2414) (Homo sapiens)	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 S. pyogenes dCas9 with c-terminal fusion of FL human p300 (aa 2-2414) driven by CMV promoter pcDNA3.1 EP300 KAT3B, RSTS2, p300
pcDNA-dCas9-p300 Core	S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens)	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 S. pyogenes dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664) driven by CMV promoter pcDNA3.1 EP300 KAT3B, RSTS2, p300
pcDNA-dCas9-p300 Core (D1399Y)	S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens)	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 S. pyogenes dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664; inactivating mutation D1399Y) driven by CMV promoter pcDNA3.1 EP300 KAT3B, RSTS2, p300
pcDNA-dCas9-p300 Core (1645/1646 RR/EE)	S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens)	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 S. pyogenes dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664; mutation 1645/1646 RR/EE) driven by CMV promoter pcDNA3.1 EP300 KAT3B, RSTS2, p300
pcDNA-dCas9-p300 Core (C1204R)	S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens)	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 S. pyogenes dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664; mutation C1204R) driven by CMV promoter pcDNA3.1 EP300 KAT3B, RSTS2, p300
pcDNA-dCas9-p300 Core (Y1467F)	S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens)	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 S. pyogenes dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664; inavtivating mutation Y1467F) driven by CMV promoter pcDNA3.1 EP300 KAT3B, RSTS2, p300
pcDNA-dCas9-p300 Core (1396/1397 SY/WW)	S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens)	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 S. pyogenes dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664; inactivating mutation 1396/1397 SY/WW) driven by CMV promoter pcDNA3.1 EP300 KAT3B, RSTS2, p300
pcDNA-dCas9-p300 Core (H1415A/E1423A/Y1424A/L1428S/Y1430A/H1434A)	S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens)	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 S. pyogenes dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664; inactivating mutations H1415A/E1423A/Y1424A/L1428S/Y1430A/H1434A) driven by CMV promoter pcDNA3.1 EP300 KAT3B, RSTS2, p300
pcDNA3.3-Nm-dCas9-p300 Core	Nm-dCas9-p300 Core (Homo sapiens)	CMV	Neomycin (select with G418)	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 N. meningiditis dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664) driven by CMV promoter pcDNA3.3 EP300 KAT3B, RSTS2, p300
dCAS9-VP64_GFP	dCAS9(D10A,H840A)-VP64_2A_GFP (Synthetic)	EF1A	GFP	Zhang	Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature. 2014 Dec 10. doi: 10.1038/nature14136.	Expresses dCAS9-VP64 activator with 2A GFP lenti(AMP)
lenti dCAS-VP64_Blast	dCAS9(D10A, N863A)-VP64_2A_Blast (Synthetic)	EF1A	Blasticidin	Zhang	Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature. 2014 Dec 10. doi: 10.1038/nature14136.	3rd generation lenti vector encoding dCAS9-VP64 with 2A Blast resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE) plenti
TetO-FUW-VdC9BV	VP64dCas9BFPVP64 (Synthetic)			Leong	A CRISPR/Cas9-Based System for Reprogramming Cell Lineage Specification. Stem Cell Reports. 2014 Dec 9;3(6):940-7. doi: 10.1016/j.stemcr.2014.09.013. Epub 2014 Oct 23.	Expresses RNA-Guided, Nuclease-Inactive VP64:dCas9-BFP:VP64—VdC9BV—Fusion Protein to Enable Transactivation of Endogenous Genes TetO-FUW
pJZC32	sgRNA, MCP-VP64	U6, CMV	Marked 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 MCP-VP64 effector for mammalian cells MP177_U6 (derived from pSico)
pJZC25	sgRNA + 1x MS2 binding module, MCP-VP64	U6, CMV	Marked 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 MS2 with MCP-VP64 effector for mammalian cells MP177_U6 (derived from pSico)
pJZC33	sgRNA + 2x MS2 binding module, MCP-VP64	U6, CMV	Marked 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 + 2x MS2 with MCP-VP64 effector for mammalian cells MP177_U6 (derived from pSico)
pJZC34	sgRNA + 2x MS2(wt+f6) binding module, MCP-VP64	U6, CMV	Marked 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 + 2x MS2(wt+f6) with MCP-VP64 effector for mammalian cells MP177_U6 (derived from pSico)
pJZC41	sgRNA, PCP-VP64	U6, CMV	Marked 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 PCP-VP64 effector for mammalian cells MP177_U6 (derived from pSico)
pJZC39	sgRNA + 1x PP7, mCherry	U6, CMV	Marked 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 PP7 with mCherry for mammalian cells MP177_U6 (derived from pSico)
pJZC101	sgRNA, COM-VP64	U6, CMV	Marked 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-VP64 effector for mammalian cells MP177_U6 (derived from pSico)
pJZC48	sgRNA + 1x COM binding module, COM-VP64	U6, CMV	Marked 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-VP64 effector for mammalian cells MP177_U6 (derived from pSico)
pJZC116	sgRNA + 2x MS2 (wt+f6) binding module, MCP-VP64	U6, CMV	Marked by BFP	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 + 2x MS2(wt+f6) with MCP-VP64 effector for mammalian cells, marked by BFP MP177_U6 (derived from pSico)
PX855	SpCas9 (aa 2-535)	CBh		Zhang	A split-Cas9 architecture for inducible genome editing and transcription modulation. Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149.	N-term SpCas9 piece of inducible transcriptional activator (dCas9(N)-FRB-2xNES) PX330
PX856	SpCas9 (aa536-1368)	CBh		Zhang	A split-Cas9 architecture for inducible genome editing and transcription modulation. Nat Biotechnol. 2015 Feb 2. doi: 10.1038/nbt.3149.	C-term SpCas9 piece of inducible transcriptional activator (dCas9(C)-FKBP-2xNLS-VP64) PX330
SP-dCas9-VPR	SP-dCas9-VPR (Homo sapiens)	CMV	Neomycin (select with G418)	Church	Highly efficient Cas9-mediated transcriptional programming. Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312.	SP-dCas9 with VP64-p65-Rta (VPR) fused to it's C-terminus; mammalian vector pcDNA3.3 TOPO
M_ST1n_VPR	ST1-dCas9-VPR (Homo sapiens)	CMV	Neomycin (select with G418)	Church	Highly efficient Cas9-mediated transcriptional programming. Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312.	ST1-dCas9 with VP64-p65-Rta (VPR) fused to it's C-terminus; mammalian vector pcDNA3.3 TOPO
PB-TRE-dCas9-VPR	dCas9-VPR (Homo sapiens)	TRE	Hygromycin	Church	Highly efficient Cas9-mediated transcriptional programming. Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312.	SP-dCas9-VPR with doxycycline-inducible expression PB-TRE
NLS-dCas9-trCIB1	dCas9-trCIB1 fusion (Synthetic)	CMV	Neomycin (select with G418)	Sato	CRISPR-Cas9-based Photoactivatable Transcription System. Chem Biol. 2015 Feb 19;22(2):169-74. doi: 10.1016/j.chembiol.2014.12.011. Epub 2015 Jan 22.	Photoactivatable transcription system. Expression of genomic anchor probe, containing dCas9 and CIB1 pcDNA3.1/V5-His A
pJZC42	sgRNA + 1XPP7, PCP-VP64 IRES mCherry	U6, CMV	mCherry fluorescence	Lim	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 + 1XPP7 with PCP-VP64 effector for mammalian cells, marked by mCherry pSico
pJZC43	sgRNA + 2XPP7, PCP-VP64 IRES mCherry	U6, CMV	mCherry fluorescence	Lim	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 + 2XPP7 with PCP-VP64 effector for mammalian cells, marked by mCherry pSico
pEF_dCas9-VP64	dCas9 (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 fused to the VP64 transcription activator, in mammalian cells, under an EF1-alpha promoter. pNEB193 Tags / Fusion Proteins 3xFLAG (C terminal on insert) VP64 (C terminal on insert)
AAV_NLS-dSaCas9-NLS-VPR	dSaCas9-VPR (Synthetic)	CMV		Church	Cas9 gRNA engineering for genome editing, activation and repression. Nat Methods. 2015 Sep 7. doi: 10.1038/nmeth.3580.	AAV vector containing nuclease null SaCas9 fused to VPR pX600-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA Plasmid #61592
CAG-DDdCas9VP192-T2A-EGFP-ires-puro	DD-dCas9VP192-T2A-EGFP (Other)	CAG	Puromycin	Otonkoski	Conditionally Stabilized dCas9 Activator for Controlling Gene Expression in Human Cell Reprogramming and Differentiation. Stem Cell Reports. 2015 Sep 8;5(3):448-59. doi: 10.1016/j.stemcr.2015.08.001.	DHFR destabilised domain (DD) fused to dCas9VP192 (S.pyogenes) on CAG expression vector. DDdCas9VP192 protein is stabilised by Trimethoprim. PyCAG
pCXLE-dCas9VP192-T2A-EGFP-shP53	dCas9-dCas9VP192-GFP-shp53 (Other)	CAG		Otonkoski	Conditionally Stabilized dCas9 Activator for Controlling Gene Expression in Human Cell Reprogramming and Differentiation. Stem Cell Reports. 2015 Sep 8;5(3):448-59. doi: 10.1016/j.stemcr.2015.08.001.	Episomal plasmid encoding dCas9VP192 and p53 shRNA pCXLE
pCXLE-dCas9VP192-T2A-EGFP	dCas9-dCas9VP192-EGFP (Other)	CAG		Otonkoski	Conditionally Stabilized dCas9 Activator for Controlling Gene Expression in Human Cell Reprogramming and Differentiation. Stem Cell Reports. 2015 Sep 8;5(3):448-59. doi: 10.1016/j.stemcr.2015.08.001.	Episomal plasmid encoding dCas9VP192 pCXLE
pHRm-NLS-dCas9-GFP11x7-NLS-P2A-BFP-NLS	NLS-dCas9-GFP11x7-NLS-P2A-BFP-NLS (Synthetic)			Huang	Versatile protein tagging in cells with split fluorescent protein. Nat Commun. 2016 Mar 18;7:11046. doi: 10.1038/ncomms11046.	Expresses NLS-dCas9-GFP11x7-NLS and BFP-NLS in mammalian cells pHRdSV40-NLS-dCas9-24xGCN4_v4-NLS-P2A-BFP-dWPRE (addgene #60910)
pAC164-pmax-dCas9Master-VP64	dCas9-VP64 (Synthetic)	CAGGS + chim intron		Cheng	Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15.	dCas9-3xGGGGS-VP64 in pmax expression vector pAC90-pmax-DEST
pAC1364-pmax-dCas9Master_mCBPHAT	dCas9-mCBPHAT (Synthetic)	CAGGS + chim intron		Cheng	Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15.	dCas9Master_mCBPHAT in pmax expression vector pAC90-pmax-DEST
pAC1410-pmax-dCas9Master_p65HSF1	dCas9-p65HSF1 (Synthetic)	CAGGS + chim intron		Cheng	Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15.	dCas9Master_p65HSF1 in pmax expression vector pAC90-pmax-DEST
lentiSAMv2		U6 and EF1A	Blasticidin	Zhang	Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. Epub 2017 Mar 23.	lenti sgRNA cloning backbone with MS2 loops at tetraloop and stemloop 2, dCas9-VP64, and blast resistance marker. Contains BsmBI sites for insertion of spacer sequences. plenti
pLV-dCas9-p300-P2A-PuroR	S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens), pac from Streptomyces alboniger (Other)	EFS	Puromycin	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-p300-P2A-PuroR lentiCRISPR v2 EP300 KAT3B, RSTS2, p300
AAVS1-idCas9-vpr	dCas9-VPR (Other)	TRE-tight	Puromycin	Na	An inducible CRISPR-ON system for controllable gene activation in human pluripotent stem cells. Protein Cell. 2017 Jan 23. doi: 10.1007/s13238-016-0360-8.	inducible CRISPR-ON system for controllable gene activation; this plasmid is used to insert dCas9-VPR casette in one allele of AAVS1 locus AAVS1 homologous recombineering donor plasmid
lentiSAM v2 (Puro)		U6 (sgRNA) and EF1a (dCas9-VP64)	Puromycin	Karpf	Lenti CRISPR Activate (unpublished)	Modified version of lentiSAM v2, a lenti sgRNA cloning backbone with MS2 loops at tetraloop/stemloop 2, dCas9-VP64, and puro resistance marker. Contains BsmBI sites for insertion of spacer sequences. lentiSAM v2
pXPR_120	dCas9 (Other)	EF1a	Blasticidin	Root	Najm et al. (unpublished)	for CRISPRa, lentiviral expression of dCas9-VPR 2A BlastR pXPR Tag / Fusion Protein VPR (C terminal on insert)

Bacteria

Plasmid	Gene/Insert	Promoter	PI	Publication	Hidden Extra Search Info
pWJ66	tracrRNA (Other), dcas9-w (Other), CRISPR array		Marraffini	Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic Acids Res. 2013 Jun 12.	Same as pdCas9, but with the dCas9-w fusion (w is fused at the C-terminal end of dCas9) pACYC184
pWJ68	tracrRNA (Other), w-dcas9 (Other), CRISPR array		Marraffini	Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic Acids Res. 2013 Jun 12.	Same as pdCas9, but with the w-dCas9 fusion (w is fused at the N-terminal end of dCas9) pACYC184
pET-dCas9-VP64-6xHis	dCas9-VP64 (Other)	T7	Liu	Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30.	Expression of dCas9-VP64-6xHis in bacterial cells pET29

Yeast

Plasmid	Gene/Insert	Promoter	Selectable Marker	PI	Publication	Hidden Extra Search Info
pTPGI_dCas9_VP64	dCas9_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)
pJZC519	dCas9-VP64 (Other)	pTdh3	LEU2	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
pJZC620	MCP-VP64, PCP-VP64, dCas9	pAdh, pAdh, pTdh3	LEU2	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
pJZC638	MCP-VP64, dCas9 (Other)	pAdh, pGal10	LEU2	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
pJZC545	sgRNA + 1x MS2 binding module	SNR52	URA3	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 MS2 for yeast cells pRS416
pJZC583	sgRNA + 2x MS2 binding module	SNR52	URA3	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 2x MS2 for yeast cells pRS416
pJZC548	sgRNA + 1x PP7	SNR52	URA3	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
pJZC603	sgRNA + 2x PP7 RNA binding module	SNR52	URA3	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 2x PP7 for yeast cells pRS416
pJZC572	sgRNA + 1x com RNA binding module	SNR52	URA3	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
pJZC593	sgRNA + MS2-PP7 RNA binding module	SNR52	URA3	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 MS2-PP7 for yeast cells pRS416
pJZC625	sgRNA +1x MS2, Pol II promoter with ribozyme cleavage	pAdh	URA3	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 MS2, Pol II promoter with ribozyme-gRNA-ribozyme design for yeast cells pSV616
pJZC595	MCP-VP64, dCas9	pAdh, Tdh3	LEU2	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-VPR	dCas9-VPR (Saccharomyces cerevisiae)	GPD	TRP1	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
pTPGI_dCas9	Yeast-optimized dCas9 (Saccharomyces cerevisiae)	pTPGI	TRP1	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-Gal4-dCas9-VP64	Gal4-dCas9-VP64 (Saccharomyces cerevisiae)	Tef1	URA3	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

Plant

Plasmid	Gene/Insert	Promoter	Selectable Marker	PI	Publication	Hidden Extra Search Info
pBUN6A11	dCas9-VP64 (Synthetic), gRNA scaffold (Synthetic)	Ubi1p, OsU3p	Bar	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-VP64, gRNA scaffold for insertion of target sequence (OsU3 promoter), Bar resistance pGreen-like binary vector
pHSN6A01	dCas9-VP64 (Synthetic), gRNA scaffold (Synthetic)	2×35Sp, AtU6-26p	Hygromycin	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-VP64, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Hyg resistance pGreen-like binary vector
pD10AH840AhCas9 (GB1041)	Cas9 coding region with mutated (D10A, H840A) and inactivated catalytic domains (human codon optimised) (Other)			Orzaez	GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. Plant Physiol. 2013 Jul;162(3):1618-31.	Provides the human codon optimized CDS of Cas9 protein with mutated (D10A, H840A) and inactivated catalytic domains as a level 0 GoldenBraid part pUPD
pYPQ152	pco-dCas9-VP64 (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-VP64 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

Drosophila

Plasmid	Gene/Insert	Promoter	PI	Publication
pAWG-dCas9-VPR	SP-dCas9-VPR (Drosophila melanogaster)	act5c	Church	Highly efficient Cas9-mediated transcriptional programming. Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312.
pWalium20-10XUAS-3XFLAG-dCas9-VPR	dCas9-VPR (Homo sapiens)	10XUAS	Perrimon	In Vivo Transcriptional Activation Using CRISPR/Cas9 in Drosophila. Genetics. 2015 Oct;201(2):433-42. doi: 10.1534/genetics.115.181065. Epub 2015 Aug 5.
pAct:dCas9-VPR	dCas9-VPR	pActin (Drosophila)	Perrimon	Comparison of Cas9 activators in multiple species. Nat Methods. 2016 May 23. doi: 10.1038/nmeth.3871.
pAct:dCas9-GCN4	dCas9-10XGCN4	pActin (Drosophila)	Perrimon	Comparison of Cas9 activators in multiple species. Nat Methods. 2016 May 23. doi: 10.1038/nmeth.3871.
pAct:dCas9-VP64	dCas9-VP64	pActin (Drosophila)	Perrimon	Comparison of Cas9 activators in multiple species. Nat Methods. 2016 May 23. doi: 10.1038/nmeth.3871.
pAct:dCas9	dCas9	pActin (Drosophila)	Perrimon	Comparison of Cas9 activators in multiple species. Nat Methods. 2016 May 23. doi: 10.1038/nmeth.3871.

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