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CRISPR Plasmids: Epigenetics

Browse CRISPR Plasmids

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CRISPR Resources

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, MKHK2, 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, MKHK2, 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, MKHK2, 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, MKHK2, 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, MKHK2, 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, MKHK2, 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, MKHK2, 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, MKHK2, 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, MKHK2, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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(+)
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)hUbCEGFP; Zeo marker is outside the LTRs and will not be packaged into virus. 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)hUbCEGFP; Zeo marker is outside the LTRs and will not be packaged into virus. 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 in vivo modulation of cis-regulatory interactions and gene expression in the chicken embryo. Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 10.1242/dev.160333. 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, KDAC3, 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]-dCas914056 (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 Enx-1, Enx1h, KMT6, mKIAA4065
    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, HESJAS, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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, HESJAS, 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
    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, KDAC3, RPD3, RPD3-2
    p-dCas9-DNMT3a-Hygro DNMT3a (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-DNMT3a fusion protein pMLM3705
    p-dCas9-TET1-Hygro TET1 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-TET1 fusion protein pMLM3705
    p-dCas9-LSD1-HygroLSD1 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-LSD1 fusion protein pMLM3705
    p-dCas9-SET7-HygroSET7 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-SET7 fusion protein pMLM3705
    p-dCas9-G9a-HygroG9a (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-G9a fusion protein pMLM3705
    p-dCas9-HDAC1-HygroHDAC1 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-HDAC1 fusion protein pMLM3705
    p-dCas9-JMJD2A-HygroJMJD2A (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-JMJD2A fusion protein pMLM3705
    p-dCas9-p300-Hygrop300 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-p300 fusion protein pMLM3705
    p-dCas9-dDNMT3a-C706S-Hygro dDNMT3a (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-dDNMT3a fusion protein pMLM3705
    p-dCas9-dTET1-H1672Y-D1674A-Hygro dTET1 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-dTET1 fusion protein pMLM3705
    p-dCas9-LSD1-K661A-Hygro dLSD1 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-dLSD1 fusion protein pMLM3705
    p-dCas9-SET7-H297G-HygrodSET7 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-dSET7 fusion protein pMLM3705
    p-dCas9-HDAC1-H141A-HygrodHDAC1 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-dHDAC1 fusion protein pMLM3705
    p-dCas9-JMJD2A-S288G-T289G-HygrodJMJD2A (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-dJMJD2A fusion protein pMLM3705
    p-dCas9-p300-S1396R-S1397R-Hygrodp300 (Homo sapiens)CMV promoterHygromycin Beck Beck lab Cas9 plasmids (unpublished) transient expression of dCas9-dp300 fusion protein pMLM3705
    pcDNA-CasE-p300-FlagEcoCasE-p300CMV Gersbach Targeted transcriptional modulation with type I CRISPR-Cas systems in human cells. Nat Biotechnol. 2019 Sep 23. pii: 10.1038/s41587-019-0235-7. doi: 10.1038/s41587-019-0235-7. Encodes E.coli CRISPR Type I-E CasE with N-terminal fusion of E tag epitope and NLS and C-terminal fusion of NLS, human p300 HAT core (aa1048-1664), and Flag tag epitope driven by CMV promoter pcDNA
    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, MKHK2, RSTS2, p300
    dSpyCas9-p300-P2A-CASANOVAdCas9-p300-P2A-CASANOVA Niopek Engineered anti-CRISPR proteins for optogenetic control of CRISPR-Cas9. Nat Methods. 2018 Oct 30. pii: 10.1038/s41592-018-0178-9. doi: 10.1038/s41592-018-0178-9. Co-expresses dSpyCas9 fused to the p300 histone acetyltransferase core domain and CASANOVA pmCherry-N1
    pPB-R1R2_EF1adCas9p300_T2A_MS2p65HSF1-IRESbsdpAdCas9-p300core-T2A-MS2-p65-HSF1 (Homo sapiens)Blasticidin Wright Pooled extracellular receptor-ligand interaction screening using CRISPR activation. Genome Biol. 2018 Nov 26;19(1):205. doi: 10.1186/s13059-018-1581-3. Expresses dCas9-p300 and MS2-p65-HSF1 fusion proteins pPB-R1R2-EM7NeoPheS
    pPB-R1R2_EF1ap300dCas9VP64_T2A_MS2p65HSF1-IRESbsdpAp300core-dCas9-VP64-T2A-MS2-p65-HSF1 (Homo sapiens)Blasticidin Wright Pooled extracellular receptor-ligand interaction screening using CRISPR activation. Genome Biol. 2018 Nov 26;19(1):205. doi: 10.1186/s13059-018-1581-3. Expresses p300-dCas9-VP64 and MS2-p65-HSF1 fusion proteins pPB-R1R2-EM7NeoPheS
    pPB-R1R2_EF1aVP64dCas9p300_T2A_MS2p65HSF1-IRESbsdpAVP64-dCas9-p300core-T2A-MS2-p65-HSF1 (Homo sapiens)Blasticidin Wright Pooled extracellular receptor-ligand interaction screening using CRISPR activation. Genome Biol. 2018 Nov 26;19(1):205. doi: 10.1186/s13059-018-1581-3. Expresses VP64-dCas9-p300 and MS2-p65-HSF1 fusion proteins pPB-R1R2-EM7NeoPheS
    pEG302-SunTag-FWAgRNA-4-22aa-TET1cdgRNA4_U6_NOS_TET1CD_2xNLS_1xHA__sfGFP_scFv_UBQ10_INSULATOR_UBQ10_dCAS9_1xHA_3xNLS_10xGCN422aa_OCS (Homo sapiens)Hygromycin Jacobsen pEG302-SunTag-FWAgRNA-4-22aa-TET1cd (unpublished) SunTag CRISPR cas9 system that targets the TET1 catalytic domain to the FWA promoter pEG302
    pEG302-SunTagng-22aaNOS_TET1CD_2xNLS_1xHA__sfGFP_scFv_UBQ10_INSULATOR_UBQ10_dCAS9_1xHA_3xNLS_10xGCN422aa_OCS (Homo sapiens)Hygromycin Jacobsen pEG302-SunTag-FWAgRNA-4-22aa-TET1cd (unpublished) Encodes a SunTag CRISPR cas9 system that does not contain a guide RNA and therefore, does not target the TET1 catalytic domain to any specific region of the genome pEG302
    Pinducer20-dCas9-SunTag-2A-BFPdCas9-SunTag-2A-BFPTRENeomycin (select with G418) Goodell Homeobox oncogene activation by pan-cancer DNA hypermethylation. Genome Biol. 2018 Aug 10;19(1):108. doi: 10.1186/s13059-018-1492-3. targeted DNA methylation tool Pinducer 20
    IF311: pMAGIC (R4-R3) NLS-Sa dCas9-NLS-LSD1Sa-dCas9/LSD1 (open) (Synthetic) Newgard Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. pMAGIC R4-R3 entry plasmid, contains 2x NLS Sa-dCas9 fused to the LSD1 H3K4me1/2 demethylase for 3 or 4-component MultiSite Gateway Pro assembly. pDONR221 P4r-P3r
  • Tags / Fusion Proteins
    • NLS (N terminal on insert)
    • NLS (C terminal on insert)
    		•
    IF405: pMAGIC (R4-R3) NLS-Sa dCas9-NLS-p300coreSa-dCas9/p300core (open) (Synthetic) Newgard Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. pMAGIC R4-R3 entry plasmid, contains 2x NLS Sa-dCas9 fused to the p300 catalytic core for 3 or 4-component MultiSite Gateway Pro assembly. pDONR221 P4r-P3r
  • Tags / Fusion Proteins
    • NLS (N terminal on insert)
    • NLS (C terminal on insert)
    		•
    IP803: pMAGIC (R4-R3) NLS-Sa dCas9-NLS-Dnmt3a3LSa-dCas9/Dnmt3a-3L (open) (Synthetic) Newgard Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. pMAGIC R4-R3 entry plasmid, contains 2x NLS Sa-dCas9 fused to the Dnmt3a-3L DNA methyltransferase for 3 or 4-component MultiSite Gateway Pro assembly. pDONR221 P4r-P3r
  • Tags / Fusion Proteins
    • NLS (N terminal on insert)
    • NLS (C terminal on insert)
    		•
    KQ403: pMAGIC (R4-R3) NLS-x dCas9(3.7)-NLS-LSD1x-dCas9(3.7)/LSD1 (open) (Synthetic) Newgard Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. pMAGIC R4-R3 entry plasmid, contains 2x NLSx-dCas9(3.7) fused to the LSD1 H3K4me1/2 demethylase for 3 or 4-component MultiSite Gateway Pro assembly. pDONR221 P4r-P3r
  • Tags / Fusion Proteins
    • NLS (N terminal on insert)
    • NLS (C terminal on insert)
    		•
    KO001: pMAGIC (R4-R3) NLS-x dCas9(3.7)-NLS-p300corex-dCas9(3.7)/p300core (open) (Synthetic) Newgard Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. pMAGIC R4-R3 entry plasmid, contains 2x NLS x-dCas9(3.7) fused to the p300 catalytic core for 3 or 4-component MultiSite Gateway Pro assembly. pDONR221 P4r-P3r
  • Tags / Fusion Proteins
    • NLS (N terminal on insert)
    • NLS (C terminal on insert)
    		•
    KO101: pMAGIC (R4-R3) NLS-x dCas9(3.7)-NLS-Dnmt3a3Lx-dCas9(3.7)/Dnmt3a-3L (open) (Synthetic) Newgard Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. pMAGIC R4-R3 entry plasmid, contains 2x NLS x-dCas9(3.7) fused to the Dnmt3a-3L DNA methyltransferase for 3 or 4-component MultiSite Gateway Pro assembly. pDONR221 P4r-P3r
  • Tags / Fusion Proteins
    • NLS (N terminal on insert)
    • NLS (C terminal on insert)
    		•
    KL202: pMAGIC (R4-R3) NLS-(SrfI/PmeI)-NLS-LSD12x NLS/LSD1 (open) (Synthetic) Newgard Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. pMAGIC R4-R3 entry plasmid, contains 2x NLS/LSD1 scaffold enabling addition of dCas9 mutants into pMAGIC. dCas9 can be inserted via unique SrfI/PmeI restriction sites pDONR221 P4r-P3r
    KL301: pMAGIC (R4-R3) NLS-(SrfI/PmeI)-NLS-p300core2x NLS/p300core (open) (Synthetic) Newgard Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. pMAGIC R4-R3 entry plasmid, contains 2x NLS/p300core scaffold enabling addition of dCas9 mutants into pMAGIC. dCas9 can be inserted via unique SrfI/PmeI restriction sites pDONR221 P4r-P3r
    KL401: pMAGIC (R4-R3) NLS-(SrfI/PmeI)-NLS-Dnmt3a3L2x NLS/Dnmt3a-3L (open) (Synthetic) Newgard Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. pMAGIC R4-R3 entry plasmid, contains 2x NLS/Dnmt3a-3L scaffold enabling addition of dCas9 mutants into pMAGIC. dCas9 can be inserted via unique SrfI/PmeI restriction sites pDONR221 P4r-P3r
    pcDNA-LmoCas6-p300LmoCas6-p300U6, CMV Gersbach Targeted transcriptional modulation with type I CRISPR-Cas systems in human cells. Nat Biotechnol. 2019 Sep 23. pii: 10.1038/s41587-019-0235-7. doi: 10.1038/s41587-019-0235-7. Encodes L.monocytogenes CRISPR Type I-B Cas6 with N-terminal fusion of Flag epitope and NLS and C-terminal fusion of NLS, human p300 HAT core (aa1048-1664), and Flag tag epitope driven by CMV promoter pcDNA
    pcDNA3.1-dCas9-Dnmt3a-Dnmt3l-P2A-eGFPdCas9m4-sv40NLS-DNMT3A-3L-sv40NLS-P2A-eGFP (Synthetic)T7 Promoter Bernstein Epigenome editing strategies for the functional annotation of CTCF insulators. Nat Commun. 2019 Sep 18;10(1):4258. doi: 10.1038/s41467-019-12166-w. Human Codon Optimized dCas9-DNMT3A3L pcDNA3.1
    pDT305dCas9-Dnmt3a-Dnmt3l-T2A-eGFP + gRNA expression cassette + LV backbone (Homo sapiens)hUBC promotereGFP Bernstein Epigenome editing strategies for the functional annotation of CTCF insulators. Nat Commun. 2019 Sep 18;10(1):4258. doi: 10.1038/s41467-019-12166-w. Human codon optimized dCas9-Dnmt3a-Dnmt3l-T2A-eGFP + gRNA expression cassette + LV backbone pDT305
    pAT888_CMV/CAG_PUFa-hTET1(CD)PUFa-hTET1CD(1418-2136) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of untagged Casilio-ME1 effector PUFa-hTET1(CD) in mammalian cells pLX302
    pAT890_CMV/CAG_dCas9-hTET1(CD)Sp dCas9-hTET1CD(1418-2136) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral plasmid for expression of Sp dCas9-hTET1(CD) fusion protein in mammalian cells pLX302
    pAT341_CMV/CAG_PUFa-TET1CD (H1671Y D1673A) (dead TET1)PUFa-dead hTET1CD(1418-2136, H1671Y D1673A) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of catalytically inactive Casilio-ME1 effector containing dead hTET1(CD) H1671Y D1673A in mammalian cells pLX302
    pAT360_CMV/CAG_PUFa-hGADD45A-hTET1(CD)PUFa-hGADD45A-hTET1CD(1418-2136) (Homo sapiens)CMV/CAG_PUFa-hGADD45A-hTET1(CD)Puromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of untagged Casilio-ME2.1 effector PUFa-hGADD45A-hTET1(CD) in mammalian cells pLX302
    pAT635_CMV/CAG_hGADD45A-PUFa-hTET1(CD)hGADD45A-PUFa-hTET1CD(1418-2136) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of untagged Casilio-ME2.2 effector hGADD45A-PUFa-hTET1(CD) in mammalian cells pLX302
    pAT608_CMV/CAG_Flag-hNEIL2-PUFa-hTET1(CD)Flag-NEILE2-PUFa-TET1(CD) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Flag-tagged Casilio-ME3.1 effector hNEIL2-PUFa-hTET1(CD) in mammalian cells pLX302
    pAT611_CMV/CAG_PUFa-hNEIL2-hTET1(CD)PUFa-NEIL2-TET1(CD) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Casilio-ME3.2 effector PUFa-hNEIL2-hTET1(CD) in mammalian cells pLX302
    pAT699-CMV/CAG_Flag-hGADD45A-PUFa-hTET1(CD)3xFlag-GADD45A-PUFa-TET1(CD) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Flag-tagged Casilio-ME2.2 effector hGADD45A-PUFa-hTET1(CD) in mammalian cells pLX302
    pAT976_CMV/CAG_Flag-hGADD45A-PUFa-dTET1CD3xFlag-GADD45A-PUFa-dTET1(CD)H1671Y D1673A) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Flag-tagged and catalytically inactive Casilio-ME2.2 effector containing dead hTET1(CD) H1671Y D1673A in mammalian cells pLX302
    pAT971_CMV/CAG_Flag-hGADD45A (G39A)-PUFa-hTET1(CD)3xFlag-GADD45A(G39A)-PUFa-TET1(CD) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Flag-tagged Casilio-ME2.2 effector with G39A GADD45A mutation hGADD45A(G39A)-PUFa-hTET1(CD) in mammalian cells pLX302
    pAT972_CMV/CAG_Flag-hGADD45A (L77E)-PUFa-hTET1(CD)3xFlag-GADD45A(L77E)-PUFa-TET1(CD) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Flag-tagged Casilio-ME2.2 effector with L77E GADD45A mutation hGADD45A (L77E)-PUFa-hTET1(CD) in mammalian cells pLX302
    pAT973_CMV/CAG_FlaghGADD45A (G39A, L77E)-PUFa-hTET1(CD)3xFlag-GADD45A(G39A L77E)-PUFa-TET1(CD) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Flag-tagged Casilio-ME2.2 effector with G39A L77E GADD45A mutations hGADD45A(G39A L77E)-PUFa-hTET1(CD) in mammalian cells pLX302
    pAT977_CMV/CAG_Flag-hNEIL2-PUFa-dTET1(CD)Flag-NEILE2-PUFa-dTET1(CD)H1671Y D1673A (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Flag-tagged Casilio-ME3.1 effector containing a catalytically dead hTET1(CD) in mammalian cells pLX302
    pAT1059_CMV/CAG_Flag-hNEIL2 (C291S)-PUFa-hTET1(CD)Flag-NEIL2(C291S)-PUFa-TET1(CD) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Flag-tagged Casilio-ME3.1 effector with C291S NEIL2 mutation hNEIL2(C291S)-PUFa-hTET1(CD) in mammalian cells pLX302
    pAT1060_CMV/CAG_Flag-hNEIL2 (R310Q)-PUFa-hTET1(CD)Flag-NEIL2(R310Q)-PUFa-TET1(CD) (Homo sapiens)CMV/CAGPuromycin Cheng Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun. 2019 Sep 20;10(1):4296. doi: 10.1038/s41467-019-12339-7. 3rd generation lentiviral expression of Flag-tagged Casilio-ME3.1 effector with R310Q NEIL2 mutation hNEIL2(R310-Q)-PUFa-hTET1(CD) in mammalian cells pLX302

    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.
    pEG302-SunTag-FWAgRNA-4-22aa-TET1cdgRNA4_U6_NOS_TET1CD_2xNLS_1xHA__sfGFP_scFv_UBQ10_INSULATOR_UBQ10_dCAS9_1xHA_3xNLS_10xGCN422aa_OCS (Homo sapiens)Hygromycin Jacobsen pEG302-SunTag-FWAgRNA-4-22aa-TET1cd (unpublished)


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