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.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 100935 | LLP109 pEF1a-gRNA-GFP | GFP, puromycin and gRNA scaffold driven by EF1a promoter |
| 100936 | LLP185 pLVP-dCas9-DNMT3a V2 | dCas9 and DNMT WT on C terminus, 4 NLS, driven by pGK promoter, with P2A site and PURO gene, within a lentiviral transfer backbone |
| 100937 | LLP245 pEF1a-TALE-DNMT3a-Wt | Empty TALE vector driven by EF1a promoter, no Puro, contains DNMT3a catalytic domain |
| 100938 | LLP249 pGK-dCas9-DNMT3a V3 | dCas9 and DNMT WT driven by pGK promoter and Puromycin driven by EF1a |
| 100939 | LLP250 pEF1a-TALE-SunTag | Empty TALE vector fused to SunTagx10 driven by EF1a promoter, with 3xHA tag, no PURO gene |
| 100940 | LLP251 pEF1a-UNC5C-4-SunTag | TALE vector with UNC5C-4 insert fused to SunTagx10, driven by EF1a, with 3xHA tag, no PURO gene |
| 100941 | LLP252 pEF1a-NLS-scFvGCN4-DNMT3a | Single chain variable fragment antibody GCN4 fused to DNMT3a, binds SunTag, with 3xTy1 tag |
| 100942 | LLP338 pEF1a-DACH1-1-SunTag | TALE vector with DACH1-1 insert fused to SunTagx10, driven by EF1a, with 3xHA tag, no PURO gene |
| 100943 | LLP339 pGK-dCas9-SunTag | dCas9 vector with SunTagx10, driven by pGK, with 3xHA tag, Puro driven by EF1a |
| 100944 | LLP340 pEF1a-NLS-scFvGCN4-DNMT3a (mutant) | Single chain variable fragment antibody GCN4 fused to DNMT3a catalytic mutant, binds SunTag, with 3xTy1 tag |
| 100945 | LLP453 pEf1a-NLS-aGCN4-mCherry | mCherry fusion to single change variable fragment antibody GCN4 for use in SunTag system , with 3xTy1 tag |
| 100957 | LLP457 pGK-dCas9-Suntag-BFP | BPF version of LLP339, Puro is replaced by BFP driven by EF1a, dCas9 with SungTagx10 driven by pGK, with 3xHA tag |
| 100958 | LLP469 pEF1a-mCherry-EMPTY-gRNA | mCherry driven by EF1a, empty gRNA scaffold driven by U6 |