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CRISPR Plasmids: Epigenetics


To make targeted epigenetic modifications, researchers have fused catalytically dead dCas9 to epigenetic modifiers. Design your gRNA to target a specific promoter or enhancer for your gene of interest.

Available modifications include:

  • histone acetylation by p300
  • histone demethylation by LSD1
  • cytosine methylation by DNMT3A or MQ1
  • cytosine demethylation by Tet1

These modifications persist over time and are potentially heritable in dividing cells. CRISPR Activate and Interfere technologies also indirectly modify local chromatin state.

CRISPR epigenetics schematic

Browse, sort, or search the tables below for CRISPR plasmids for epigenetic modification in mammalian or plant systems.

Mammalian

Plasmid Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
pcDNA-dCas9-FLp300S.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 CoreS.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens)CMVNeomycin (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 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 CoreNm-dCas9-p300 Core (Homo sapiens)CMVNeomycin (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
pCCL-PGK-SPdCas9-BFP-DNMT3AdSpCas9-BFP-DNMT3A, catalytic domain of DNMT3A (Synthetic) Luo Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. dCas9 fused to BFP and the human DNMT3A catalytic domain 3rd Generation lentiviral vector
pCCL-PGK-SPdCas9-BFP-DNMT3BdSpCas9-BFP-DNMT3B, catalytic domain of DNMT3B (Synthetic) Luo Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. dCas9 fused to BFP and the human DNMT3B catalytic domain 3rd Generation lentiviral vector
pCCL-PGK-SPdCas9-BFP-DNMT3A(E752A)dSpCas9-BFP-DNMT3A(E752A), DNMT3A catalytic domain with inactivation mutation E752A (Synthetic) Luo Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. dCas9 fused to BFP and the human DNMT3A catalytically inactive domain 3rd Generation lentiviral vector
pCCL-PGK-SPdCas9-BFP-DNMT3B(E697A)dSpCas9-BFP-DNMT3B(E697A), DNMT3B catalytic domain with inactivation mutation E697A (Other) Luo Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. dCas9 fused to BFP and the human DNMT3B catalytically inactive domain 3rd Generation lentiviral vector
pLenti-EF1a-SPdCas9-DNMT3A-2A-BlastdSpCas9-DNMT3A catalytic domain (Synthetic) Luo Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. dCas9 fused to human DNMT3A catalytic domain 3rd generation lentiviral vector
pLenti-EF1a-SPdCas9-DNMT3B-2A-BlastdSpCas9-DNMT3B catalytic domain (Synthetic) Luo Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. dCas9 fused to human DNMT3B catalytic domain 3rd generation lentiviral vector
pLenti-EF1a-SPdCas9-DNMT3A(E752A)-2A-BlastdSpCas9-DNMT3A(E752A) Luo Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. dCas9 fused to the human DNMT3A catalytically inactive domain 3rd generation lentiviral vector
pLenti-EF1a-SPdCas9-DNMT3B(E697A)-2A-BlastdSpCas9-DNMT3B(E697A), DNMT3B catalytic domain with inactivation mutation E697A (Other) Luo Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. dCas9 fused to the human DNMT3B catalytically inactive domain 3rd Generation lentiviral vector
pdCas9-DNMT3A-EGFPS. pyogenes dCas9 fused with the catalytic domain of human DNMT3A (amino acids P602-V912) and T2A-EGFP (Homo sapiens)CBh Zoldoš Repurposing the CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Res. 2016 Mar 11. pii: gkw159. Expression of human codon-optimized dCas9-DNMT3A fusion with T2A-EGFP for targeted DNA methylation in mammalian cells; cloning backbone for sgRNA PX462 DNMT3A DNMT3A2, M.HsaIIIA, TBRS
pdCas9-DNMT3A-PuroRS. pyogenes dCas9 fused with the catalytic domain of human DNMT3A (amino acids P602-V912) and T2A-PuroR (Homo sapiens)CBh Zoldoš Repurposing the CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Res. 2016 Mar 11. pii: gkw159. Expression of human codon-optimized dCas9-DNMT3A fusion with T2A-PuroR for targeted DNA methylation in mammalian cells; cloning backbone for sgRNA PX462 DNMT3A DNMT3A2, M.HsaIIIA, TBRS
pdCas9-DNMT3A-PuroR (ANV)S. pyogenes dCas9 fused with the inactivated (E756A) catalytic domain of human DNMT3A (amino acids P602-V912) and T2A-PuroR (Homo sapiens)CBh Zoldoš Repurposing the CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Res. 2016 Mar 11. pii: gkw159. Expression of mutant human codon-optimized dCas9-DNMT3A fusion (inactive catalytic domain of DNMT3A) with T2A-PuroR; cloning backbone for sgRNA PX462 DNMT3A DNMT3A2, M.HsaIIIA, TBRS
pdCas9-DNMT3A-EGFP (ANV)S. pyogenes dCas9 fused with the inactivated (E756A) catalytic domain of human DNMT3A (amino acids P602-V912) and T2A-EGFP (Homo sapiens)CBh Zoldoš Repurposing the CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Res. 2016 Mar 11. pii: gkw159. Expression of mutant human codon-optimized dCas9-DNMT3A fusion (inactive catalytic domain of DNMT3A) with T2A-EGFP; cloning backbone for sgRNA PX462 DNMT3A DNMT3A2, M.HsaIIIA, TBRS
pdCas9-DNMT3A-PuroR_hNTNon-targeting sgRNA human (Homo sapiens)U6 Zoldoš Repurposing the CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Res. 2016 Mar 11. pii: gkw159. Expression of dCas9-DNMT3A fusion with T2A-PuroR and non-targeting control sgRNA for use in human cells pdCas9-DNMT3A-PuroR
pdCas9-DNMT3A-PuroR_v2S. pyogenes dCas9 fused with the catalytic domain of human DNMT3A (amino acids P602-V912) and T2A-PuroR_v2 (Homo sapiens)CBhPuromycin Zoldoš Repurposing the CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Res. 2016 Mar 11. pii: gkw159. Expression of human codon-optimized dCas9-DNMT3A fusion with T2A-PuroR (v2) for targeted DNA methylation in mammalian cells; cloning backbone for sgRNA PX462 DNMT3A DNMT3A2, M.HsaIIIA, TBRS
pdCas9-DNMT3A-PuroR (ANV)_v2S. pyogenes dCas9 fused with the inactivated (E756A) catalytic domain of human DNMT3A (amino acids P602-V912) and T2A-PuroR (v2) (Homo sapiens)CBhPuromycin Zoldoš Repurposing the CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Res. 2016 Mar 11. pii: gkw159. Expression of mutant human codon-optimized dCas9-DNMT3A fusion (inactive catalytic domain of DNMT3A) with T2A-PuroR (v2); cloning backbone for sgRNA PX462 DNMT3A DNMT3A2, M.HsaIIIA, TBRS
TetO-dCas9-D3ADNMT3A CD aa 598-912 (Homo sapiens)CMV Challen Reprogrammable CRISPR/Cas9-based system for inducing site-specific DNA methylation. Biol Open. 2016 May 11. pii: bio.019067. doi: 10.1242/bio.019067. Expresses dead Cas9 (dCas9) fused to DNMT3A catalytic domain (CD) linked to IRES-mCherry under the control of TetOp and a minimal CMV promoter Addgene # 20321
TetO-dCas9-mD3ADNMT3A CD aa 598-912 (Homo sapiens)CMV Challen Reprogrammable CRISPR/Cas9-based system for inducing site-specific DNA methylation. Biol Open. 2016 May 11. pii: bio.019067. doi: 10.1242/bio.019067. Expresses dead Cas9 (dCas9) fused to inactive DNMT3A catalytic domain (CD) linked to IRES-mCherry under the control of TetOp and a minimal CMV promoter Addgene # 20321 DNMT3A DNMT3A2, M.HsaIIIA, TBRS
pCMV-dCas9-D3ADNMT3A CD aa 598-912 (Homo sapiens)pCMV Challen Reprogrammable CRISPR/Cas9-based system for inducing site-specific DNA methylation. Biol Open. 2016 May 11. pii: bio.019067. doi: 10.1242/bio.019067. Expresses dead Cas9 (dCas9) fused to DNMT3A catalytic domain (CD) under the control of the CMV promoter pCMV DNMT3A DNMT3A2, M.HsaIIIA, TBRS
pCMV-dCas9-mD3ADNMT3A CD aa 598-912 (Homo sapiens)pCMV Challen Reprogrammable CRISPR/Cas9-based system for inducing site-specific DNA methylation. Biol Open. 2016 May 11. pii: bio.019067. doi: 10.1242/bio.019067. Expresses dead Cas9 (dCas9) fused to inactive DNMT3A catalytic domain (CD) under the control of the CMV promoter pCMV DNMT3A DNMT3A2, M.HsaIIIA, TBRS
pPlatTET-gRNA2dCas9-5xPlat2AflD-P2A-scFvGCN4sfGFPTET1CDNeomycin (select with G418) Hatada Targeted DNA demethylation in vivo using dCas9-peptide repeat and scFv-TET1 catalytic domain fusions. Nat Biotechnol. 2016 Aug 29. doi: 10.1038/nbt.3658. All in one vector which contains Cas9 peptide array (linker length: 22aa), antibody-sfGFP-TET1CD, and gRNA expression system. pCAG
pCAG-scFvGCN4sfGFPTET1CDscFvGCN4sfGFPTET1CDNeomycin (select with G418) Hatada Targeted DNA demethylation in vivo using dCas9-peptide repeat and scFv-TET1 catalytic domain fusions. Nat Biotechnol. 2016 Aug 29. doi: 10.1038/nbt.3658. Expression vector for antibody-sfGFP-TET1CD. pCAG
pdCas9-Tet1-CDdCas9-Tet1-CD and sgRNA scaffold with 2xMS2 binding sites (Synthetic)U6, CBH Hu A CRISPR-based approach for targeted DNA demethylation. Cell Discov. 2016 May 3;2:16009. doi: 10.1038/celldisc.2016.9. eCollection 2016. To introduce dCas9-fused Tet1-CD and sgRNA2.0 system pX330
pcDNA3.1-MS2-Tet1-CDMS2-Tet1-CD (Synthetic)CMVHygromycin Hu A CRISPR-based approach for targeted DNA demethylation. Cell Discov. 2016 May 3;2:16009. doi: 10.1038/celldisc.2016.9. eCollection 2016. To introduce MS2 coat protein fused Tet1-CD pcDNA3.1-hygro(+)
pLV-dCas9-p300-P2A-PuroRS.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) (Homo sapiens), pac from Streptomyces alboniger (Other)EFSPuromycin 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
Fuw-dCas9-Tet1CDdCas9-Tet1CD (Homo sapiens)Zeo marker is outside the LTRs and will not be packaged into virus Jaenisch Editing DNA Methylation in the Mammalian Genome. Cell. 2016 Sep 22;167(1):233-247.e17. doi: 10.1016/j.cell.2016.08.056. Lentiviral construct to express dCas9 fused with the catalytic domain of Tet1 FUW
Fuw-dCas9-Dnmt3adCas9-Dnmt3a (Homo sapiens)Zeocin ; Resistance marker is outside the LTRs and will not be packaged into virus Jaenisch Editing DNA Methylation in the Mammalian Genome. Cell. 2016 Sep 22;167(1):233-247.e17. doi: 10.1016/j.cell.2016.08.056. Lentiviral construct to express dCas9 fused with Dnmt3a FUW
Fuw-dCas9-Dnmt3a_IMdCas9-Dnmt3a_IM (Homo sapiens)Zeocin ; Resistance marker is outside the LTRs and will not be packaged into virus Jaenisch Editing DNA Methylation in the Mammalian Genome. Cell. 2016 Sep 22;167(1):233-247.e17. doi: 10.1016/j.cell.2016.08.056. Lentiviral construct to express dCas9 fused with an inactive Dnmt3a catalytic domain (E664A, E756A) FUW
Fuw-dCas9-Tet1CD_IMdCas9-Tet1CD_IM (Homo sapiens)Zeocin ; Resistance marker is outside the LTRs and will not be packaged into virus Jaenisch Editing DNA Methylation in the Mammalian Genome. Cell. 2016 Sep 22;167(1):233-247.e17. doi: 10.1016/j.cell.2016.08.056. Lentiviral construct to express dCas9 fused with an inactive catalytic domain of Tet1 (H1672Y, D1674A) FUW
Fuw-dCas9-Dnmt3a-P2A-tagBFPdCas9-Dnmt3a (Homo sapiens) Jaenisch Editing DNA Methylation in the Mammalian Genome. Cell. 2016 Sep 22;167(1):233-247.e17. doi: 10.1016/j.cell.2016.08.056. Lentiviral construct to express dCas9 fused with Dnmt3a-P2A-tagBFP FUW
138-dCas9-Dnmt3adCas9-Dnmt3a (Homo sapiens)Neomycin (select with G418) Jaenisch Editing DNA Methylation in the Mammalian Genome. Cell. 2016 Sep 22;167(1):233-247.e17. doi: 10.1016/j.cell.2016.08.056. PiggyBac transposon system construct with dCas9-Dnmt3a PiggyBac
pcDNA3.1-dCas9-MQ1(WT)-EGFPsite-specific DNA-methyltransferase SssICMVNeomycin (select with G418) ; EGFP Goodell Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026. Targeted CpG methylation pcDNA3.1
pcDNA3.1-dCas9-MQ1(Q147L)-EGFPsite-specific DNA-methyltransferase SssI (Synthetic)CMVNeomycin (select with G418) ; EGFP Goodell Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026. Targeted CpG methylation pcDNA3.1
pcDNA3.1-dCas9-MQ1(C141S)-EGFPsite-specific DNA-methyltransferase SssI (Synthetic)CMVNeomycin (select with G418) ; EGFP Goodell Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026. Targeted CpG methylation pcDNA3.1
pcDNA3.1-dCas9-MQ1(S317A)-EGFPsite-specific DNA-methyltransferase SssI (Synthetic)CMVNeomycin (select with G418) ; EGFP Goodell Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026. Targeted CpG methylation pcDNA3.1
pcDNA3.1-dCas9-dMQ1-EGFPsite-specific DNA-methyltransferase SssI (Synthetic)CMVNeomycin (select with G418) ; EGFP Goodell Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026. Targeted CpG methylation pcDNA3.1
pLV hUbC-dCas9-MQ1(Q147L)-EGFPsite-specific DNA-methyltransferase SssI (Synthetic)hUbCZeocin ; EGFP Goodell Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026. Targeted CpG methylation pLV hUbC-dCas9-T2A-GFP (#53191)
pLV hUbC-dCas9-dMQ1-EGFGsite-specific DNA-methyltransferase SssI (Synthetic)hUbCZeocin ; EGFP Goodell Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026. Targeted CpG methylation pLV hUbC-dCas9-T2A-GFP (#53191)
pX330a dCas9-LSD1dCas9-LSD1 (Other)CAG Sauka-Spengler Genome and epigenome engineering CRISPR toolkit for probing in vivo cis-regulatory interactions in the chicken embryo BioRxiv Modified CAG promoter-containing vector for ubiquitous expression of catalytically inactive Cas9 fused to LSD1. For targeted enhancer demethylation in chicken embryos. x330
pXPR_123dCas9 (Other)EF1aBlasticidin Root Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens. Nat Biotechnol. 2018 Feb;36(2):179-189. doi: 10.1038/nbt.4048. Epub 2017 Dec 18. for CRISPRi, lentiviral expression of dCas9-p300 2A BlastR. pXPR
  • Tag / Fusion Protein
    • p300 (C terminal on insert)
  • dCas9-hHDAC3dCAS9-HDAC3 (Homo sapiens)EF1A Zhou Locus-specific histone deacetylation using a synthetic CRISPR-Cas9-based HDAC. Nat Commun. 2017 May 12;8:15315. doi: 10.1038/ncomms15315. 3rd generation lenti vector encoding dCas9-hHDAC3 (EF1a-NLS-dCas9(N863)-hHDAC3). Expresses dCas9 fused to human HDAC3. plenti HDAC3 HD3, RPD3, RPD3-2
    dCas9-FOG1[N+C]FOG1(aa 1-45) (Homo sapiens)CMVNeomycin (select with G418) Segal dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression. Nucleic Acids Res. 2017 Sep 29;45(17):9901-9916. doi: 10.1093/nar/gkx578. dCas9 fused to two FOG1 peptide [aa 1-45] , one at the N-terminus and one at the C-terminus of dCas9; pCDNA3 vector backbone, mammalian expression pcDNA3.3-TOPO ZFPM1 FOG, FOG1, ZC2HC11A, ZNF408, ZNF89A
    Ezh2[FL]-dCas9Ezh2 (Mus musculus)CMVNeomycin (select with G418) Segal dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression. Nucleic Acids Res. 2017 Sep 29;45(17):9901-9916. doi: 10.1093/nar/gkx578. Full-length [FL] mouse Ezh2 fused to N-terminus of dCas9; pCDNA3 vector backbone, mammalian expression pcDNA3.3-TOPO
    Ezh2[SET]-dCas9Ezh2[SET] (Mus musculus)CMVNeomycin (select with G418) Segal dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression. Nucleic Acids Res. 2017 Sep 29;45(17):9901-9916. doi: 10.1093/nar/gkx578. Catalytic domain [SET] of mouse Ezh2 fused to N-terminus of dCas9; pCDNA3 vector backbone, mammalian expression pcDNA3.3-TOPO
    SUV[SET]-dCas9SUVAR39H1 (Homo sapiens)CMVNeomycin (select with G418) Segal dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression. Nucleic Acids Res. 2017 Sep 29;45(17):9901-9916. doi: 10.1093/nar/gkx578. Catalytic domain [SET] of human SUVAR39H1 fused to N-terminus of dCas9; pCDNA3 vector backbone, mammalian expression pcDNA3.3-TOPO SUV39H1 H3-K9-HMTase 1, KMT1A, MG44, SUV39H
    G9A[SET]-dCas9G9A (Homo sapiens)CMVNeomycin (select with G418) Segal dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression. Nucleic Acids Res. 2017 Sep 29;45(17):9901-9916. doi: 10.1093/nar/gkx578. Catalytic domain [SET] of human G9A fused to N-terminus of dCas9; pCDNA3 vector backbone, mammalian expression pcDNA3.3-TOPO EHMT2 BAT8, C6orf30, G9A, GAT8, KMT1C, NG36
    DNMT3A-dCas9DNMT3A (Homo sapiens)CMVNeomycin (select with G418) Segal dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression. Nucleic Acids Res. 2017 Sep 29;45(17):9901-9916. doi: 10.1093/nar/gkx578. DNMT3A fused to N-terminus of dCas9; pCDNA3 vector backbone, mammalian expression pcDNA3.3-TOPO DNMT3A DNMT3A2, M.HsaIIIA, TBRS
    LLP185 pLVP-dCas9-DNMT3a V2dCas9, DNMT3a catalytic domain (Homo sapiens)Puromycin Lister A modular dCas9-SunTag DNMT3A epigenome editing system overcomes pervasive off-target activity of direct fusion dCas9-DNMT3A constructs. Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117. dCas9 and DNMT WT on C terminus, 4 NLS, driven by pGK promoter, with P2A site and PURO gene, within a lentiviral transfer backbone pLVP DNMT3A DNMT3A2, M.HsaIIIA, TBRS
    LLP249 pGK-dCas9-DNMT3a V3dCas9, DNMT3a catalytic domain (Homo sapiens)Puromycin Lister A modular dCas9-SunTag DNMT3A epigenome editing system overcomes pervasive off-target activity of direct fusion dCas9-DNMT3A constructs. Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117. dCas9 and DNMT WT driven by pGK promoter and Puromycin driven by EF1a LLP185 pLVP-dCas9-DNMT3a V2 DNMT3A DNMT3A2, M.HsaIIIA, TBRS
    LLP252 pEF1a-NLS-scFvGCN4-DNMT3ascFvGCN4, sfGFP, GB1, DNMT3a catalytic domain (Homo sapiens) Lister A modular dCas9-SunTag DNMT3A epigenome editing system overcomes pervasive off-target activity of direct fusion dCas9-DNMT3A constructs. Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117. Single chain variable fragment antibody GCN4 fused to DNMT3a, binds SunTag, with 3xTy1 tag LLP245 pEF1a-TALE-DNMT3a-Wt DNMT3A DNMT3A2, M.HsaIIIA, TBRS
    LLP340 pEF1a-NLS-scFvGCN4-DNMT3a (mutant)scFvGCN4, sfGFP, GB1, DNMT3a catalytic mutant (Homo sapiens) Lister A modular dCas9-SunTag DNMT3A epigenome editing system overcomes pervasive off-target activity of direct fusion dCas9-DNMT3A constructs. Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117. Single chain variable fragment antibody GCN4 fused to DNMT3a catalytic mutant, binds SunTag, with 3xTy1 tag LLP252 pEF1a-NLS-scFvGCN4-DNMT3a DNMT3A DNMT3A2, M.HsaIIIA, TBRS
    pINDUCER dCas9-TET1CDMutdCas9-TET1CD Mut (Homo sapiens)EGFP Huangfu TET proteins safeguard bivalent promoters from de novo methylation in human embryonic stem cells. Nat Genet. 2018 Jan;50(1):83-95. doi: 10.1038/s41588-017-0002-y. Epub 2017 Dec 4. The dCas9-TET1CDMut fusion protein is cloned into the pINDUCER lentiviral backbone (Addgene plasmid# 46948). The TET1 catalytic domain is mutated to abolish catalytic function. pINDUCER21 (ORF-EG)
    pINDUCER dCas9-TET1CDdCas9-TET1CD (Homo sapiens)EGFP Huangfu TET proteins safeguard bivalent promoters from de novo methylation in human embryonic stem cells. Nat Genet. 2018 Jan;50(1):83-95. doi: 10.1038/s41588-017-0002-y. Epub 2017 Dec 4. The dCas9-TET1CD fusion protein is cloned into the pINDUCER lentiviral backbone (Addgene plasmid# 46948). pINDUCER21 (ORF-EG)
    scFv-sfGFP-DNMT3A1DNMT3A1 (Homo sapiens) Goodell DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A. Genome Biol. 2017 Sep 18;18(1):176. doi: 10.1186/s13059-017-1306-z. The plasmid encodes a single-chain antibody that binds to the GCN4 peptide from the SunTag system, and is fused to a DNA methyltransferase DNMT3A1 pHR DNMT3A DNMT3A2, M.HsaIIIA, TBRS
    PB-CAG-DDdCas9VPP300-T2A-GFP-IRES-NeoDDdCas9VP192-P300-T2A-GFP-IRES-Neo (Other)CAGNeomycin (select with G418) Otonkoski Human pluripotent reprogramming with CRISPR activators. Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x. PiggyBac transposon system construct with constitutive CAG promoter expression of TMP inducible DDdCas9VP192-P300core activator followed by T2A-EGFP as a reporter and IRES-Neo as selection PB-CAG
    PB-CAG-DDdCas9VPPH-T2A-GFP-IRES-NeoDDdCas9VP192-p65-HSF1-P300-T2A-GFP-IRES-Neo (Other)CAGNeomycin (select with G418) Otonkoski Human pluripotent reprogramming with CRISPR activators. Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x. PiggyBac transposon system construct with constitutive CAG promoter expression of TMP inducible DDdCas9VP192-P300core-p65-HSF1 activator followed by T2A-EGFP as a reporter and IRES-Neo as selection PB-CAG
    PB-tight-DDdCas9VPP300-T2A-GFP-IRES-NeoDDdCas9VP192-P300-T2A-GFP-IRES-Neo (Other)TRE-tightNeomycin (select with G418) Otonkoski Human pluripotent reprogramming with CRISPR activators. Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x. PiggyBac construct with doxycyline inducible TRE-tight promoter expression of TMP inducible DDdCas9VP192-P300core activator followed by T2A-EGFP as a reporter and IRES-Neo as selection PB-tight
    PB-tight-DDdCas9VPPH-T2A-GFP-IRES-NeoDDdCas9VP192-p65-HSF1-P300-T2A-GFP-IRES-Neo (Other)TRE-tightNeomycin (select with G418) Otonkoski Human pluripotent reprogramming with CRISPR activators. Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x. PiggyBac construct with doxycyline inducible TRE-tight promoter expression of TMP inducible DDdCas9VP192-P300core-p65-HSF1 activator followed by T2A-EGFP as a reporter and IRES-Neo as selection PB-tight
    pCXLE-dCas9VPP300-T2A-GFP-shP53dCas9VPP300-T2A-GFP-shP53 (Other)CAG Otonkoski Human pluripotent reprogramming with CRISPR activators. Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x. EBNA episome plasmid for CAG promoter constitutive expression of dCas9-VP192-P300core followed by T2A-GFP as a reporter. Includes p53 shRNA expression cassette. pCXLE
    pCXLE-dCas9VPPH-T2A-GFP-shP53dCas9VPPH-T2A-GFP-shP53 (Other)CAG Otonkoski Human pluripotent reprogramming with CRISPR activators. Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x. EBNA episome plasmid for CAG promoter constitutive expression of dCas9-VP192-P300core-p65-HSF1 followed by T2A-GFP as a reporter. Includes p53 shRNA expression cassette. pCXLE
    pCAG-GBP-dCas9-mRFPdCas9 (Synthetic)Blasticidin Bultmann Site-specific recruitment of epigenetic factors with a modular CRISPR/Cas system. Nucleus. 2017 May 4;8(3):279-286. doi: 10.1080/19491034.2017.1292194. Epub 2017 Feb 23. Expression construct for dCas9 fused to GFP-binding Protein (N-terminally) and mRFP (C-terminus) pCAG
    dCas-hHDAC3-R265PdCas9-HDAC3-R265P (Homo sapiens)EF1A Zhou Locus-specific histone deacetylation using a synthetic CRISPR-Cas9-based HDAC. Nat Commun. 2017 May 12;8:15315. doi: 10.1038/ncomms15315. Expresses dCas9 fused to human HDAC3 with R265P mutation plenti HDAC3 HD3, RPD3, RPD3-2
    Fuw-dCas9-Tet1CD-P2A-BFPdCas9-Tet1CD-HA-P2A-BFP (Synthetic)UbcZeo marker is outside the LTRs and will not be packaged into virus Jaenisch Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene. Cell. 2018 Feb 8. pii: S0092-8674(18)30049-7. doi: 10.1016/j.cell.2018.01.012. Lentiviral vector to express dCas9-Tet1CD-P2A-tagBFP Fuw
    Fuw-dCas9-dead Tet1CD-P2A-BFPdCas9-dead Tet1CD-HA-P2A-BFP (Synthetic)UbcZeo marker is outside the LTRs and will not be packaged into virus Jaenisch Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene. Cell. 2018 Feb 8. pii: S0092-8674(18)30049-7. doi: 10.1016/j.cell.2018.01.012. Lentiviral vector to express dCas9-dead Tet1CD-P2A-tagBFP Fuw
    Pblu-AAVS1-Cas9-p300-M2rtTA-AAVS1spCas9-p300 (Homo sapiens) Rong iKA-CRISPR hESCs for inducible and multiplex orthogonal gene knockout and activation. FEBS Lett. 2018 Jun 5. doi: 10.1002/1873-3468.13127. Donor plasmid with a Cas9-p300 expression cassette, a M2rtTA expression cassette, a T2A-puro selection cassette, and two gRNA recognition sequences at both ends of the whole insertion DNA fragment. PbluSKP EP300 KAT3B, RSTS2, p300

    Plant

    Plasmid Gene/Insert Promoter Selectable Marker PI Publication
    SunTagng22aaNOS_TET1CD_2xNLS_1xHA__sfGFP_scFv_UBQ10_INSULATOR_UBQ10_dCAS9_1xHA_3xNLS_10xGCN422aa_OCS (Homo sapiens)Hygromycin Jacobsen Targeted DNA demethylation of the Arabidopsis genome using the human TET1 catalytic domain. Proc Natl Acad Sci U S A. 2018 Feb 27;115(9):E2125-E2134. doi: 10.1073/pnas.1716945115. Epub 2018 Feb 14.
    SunTagng14aaNOS_TET1CD_2xNLS_1xHA__sfGFP_scFv_UBQ10_INSULATOR_UBQ10_dCAS9_1xHA_3xNLS_10xGCN414aa_OCS (Homo sapiens)Hygromycin Jacobsen Targeted DNA demethylation of the Arabidopsis genome using the human TET1 catalytic domain. Proc Natl Acad Sci U S A. 2018 Feb 27;115(9):E2125-E2134. doi: 10.1073/pnas.1716945115. Epub 2018 Feb 14.


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