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A modular dCas9-SunTag DNMT3A epigenome editing system overcomes pervasive off-target activity of direct fusion dCas9-DNMT3A constructs.
Pflueger C, Tan D, Swain T, Nguyen TV, Pflueger J, Nefzger C, Polo JM, Ford E, Lister R
Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
100935LLP109 pEF1a-gRNA-GFPGFP, puromycin and gRNA scaffold driven by EF1a promoter
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100936LLP185 pLVP-dCas9-DNMT3a V2dCas9 and DNMT WT on C terminus, 4 NLS, driven by pGK promoter, with P2A site and PURO gene, within a lentiviral transfer backbone
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100937LLP245 pEF1a-TALE-DNMT3a-WtEmpty TALE vector driven by EF1a promoter, no Puro, contains DNMT3a catalytic domain
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100938LLP249 pGK-dCas9-DNMT3a V3dCas9 and DNMT WT driven by pGK promoter and Puromycin driven by EF1a
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100939LLP250 pEF1a-TALE-SunTagEmpty TALE vector fused to SunTagx10 driven by EF1a promoter, with 3xHA tag, no PURO gene
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100940LLP251 pEF1a-UNC5C-4-SunTagTALE vector with UNC5C-4 insert fused to SunTagx10, driven by EF1a, with 3xHA tag, no PURO gene
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100941LLP252 pEF1a-NLS-scFvGCN4-DNMT3aSingle chain variable fragment antibody GCN4 fused to DNMT3a, binds SunTag, with 3xTy1 tag
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100942LLP338 pEF1a-DACH1-1-SunTagTALE vector with DACH1-1 insert fused to SunTagx10, driven by EF1a, with 3xHA tag, no PURO gene
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100943LLP339 pGK-dCas9-SunTagdCas9 vector with SunTagx10, driven by pGK, with 3xHA tag, Puro driven by EF1a
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100944LLP340 pEF1a-NLS-scFvGCN4-DNMT3a (mutant)Single chain variable fragment antibody GCN4 fused to DNMT3a catalytic mutant, binds SunTag, with 3xTy1 tag
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100945LLP453 pEf1a-NLS-aGCN4-mCherrymCherry fusion to single change variable fragment antibody GCN4 for use in SunTag system , with 3xTy1 tag
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100957LLP457 pGK-dCas9-Suntag-BFPBPF version of LLP339, Puro is replaced by BFP driven by EF1a, dCas9 with SungTagx10 driven by pGK, with 3xHA tag
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100958LLP469 pEF1a-mCherry-EMPTY-gRNAmCherry driven by EF1a, empty gRNA scaffold driven by U6
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