...Methyltransferase 3 Alpha (DNMT3A) Vlatka Zoldoš’ lab has deposited pdCas9-DNMT3A-EGFP and pdCas9-DNMT3A-PuroR for targeted...including pcDNA-dCas9-p300 Core and pcDNA3.3-Nm-dCas9-p300 Core, as well as pLV-dCas9-p300-P2A-PuroR for...pCMV-dCas9-D3A) and Tet-dependent (TetO-dCas9-D3A) constructs. For lentiviral expression, Fuw-dCas9-Dnmt3a...Dnmt3a and Fuw-dCas9-Dnmt3a-P2A-tagBFP are available from Rudolf Jaenisch’s lab, with the former also available...as ready-to-use lentivirus. Figure 3: dCas9-DNMT3A methylates cytosines at promoters and enhancers...Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase...Fusing dCas9 to a transcriptional activator like VP64 or VPR activates transcription, whereas dCas9-KRAB...

... Tethering dCas9 to the histone acetyltransferase p300 or the DNA methyltransferase DNMT3A allows researchers...Enzymatic domains tethered to dCas9 often interact with DNA independently of dCas9 targeting (Galonska 2018...binding is crucial for dCas9 to interfere with DNMT1 even through replication. The dCas9 also needs to be removeable...has several great options: pLV hUbC-dCas9-T2A-GFP for removable dCas9 expression and pLM-CMV-R-Cre. Selection...absent before. But other enzymatic fusions, such as dCas9-TET, can, like the iron ball-and-chain that they...-based epigenetic approach (spoiler alert: it’s dCas9… with no modifications). Ready to get started?   ...dioxygenase  When I started to work on building a CRISPR/dCas9-based tool for DNA demethylation in 2015, none existed...

...protein (dCas9 shown here). D3L recruits the endogenous, autoinhibited methyltransferase DNMT3A to the ... a programmable epigenetic editor consisting of dCas9 fused to a DNA methyltransferase — that can drive...DNA-targeting proteins, so the team created versions using dCas9, zinc finger proteins, and transcription activator-like...

...catalytically inactive Cas9 (dCas9) with the catalytic domain from DNA methyltransferase, DNMT3A. Co-expression of...creating a new dCas9-VPR expression plasmid that uses the piggybac transposase to integrate dCas9-VPR into ... is genomic integration of dCas9-VPR and controlled expression of the dCas9-VPR transgene by doxycycline... activator developed by the Church lab, termed “dCas9-VPR”, which facilitates robust activation of target...target genes (20-40x greater than standard dCas9-VP64 activators) when introduced into mammalian cells....activate target genes, the relatively large size of dCas9-VPR limits one’s ability to package it into lentivirus...transfection precluding the need for virus. Furthermore, dCAS9-VPR expression is under the control of the TRE promoter...