An enhanced CRISPR repressor for targeted mammalian gene regulation.
Nan Cher YeoAlejandro ChavezAlissa Lance-ByrneYingleong ChanDavid MennDenitsa MilanovaChih-Chung KuoXiaoge GuoSumana SharmaAngela TungRyan J CecchiMarcelle TuttleSwechchha PradhanElaine T LimNoah DavidsohnMo R EbrahimkhaniJames J CollinsNathan E LewisSamira KianiGeorge M ChurchPublished in: Nature methods (2018)
The RNA-guided endonuclease Cas9 can be converted into a programmable transcriptional repressor, but inefficiencies in target-gene silencing have limited its utility. Here we describe an improved Cas9 repressor based on the C-terminal fusion of a rationally designed bipartite repressor domain, KRAB-MeCP2, to nuclease-dead Cas9. We demonstrate the system's superiority in silencing coding and noncoding genes, simultaneously repressing a series of target genes, improving the results of single and dual guide RNA library screens, and enabling new architectures of synthetic genetic circuits.