An enhanced CRISPR repressor for targeted mammalian gene regulation.
Yeo NC., Chavez A., Lance-Byrne A., Chan Y., Menn D., Milanova D., Kuo C-C., Guo X., Sharma S., Tung A., Cecchi RJ., Tuttle M., Pradhan S., Lim ET., Davidsohn N., Ebrahimkhani MR., Collins JJ., Lewis NE., Kiani S., Church GM., Church GM.
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.