An enhanced CRISPR repressor for targeted mammalian gene regulation.

Yeo, Nan Cher; Chavez, Alejandro; Lance-Byrne, Alissa; Chan, Yingleong; Menn, David; Milanova, Denitsa; Kuo, Chih-Chung; Guo, Xiaoge et al. · Nat Methods · 2018

basic_science · Level V

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Abstract

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.

Medical subject headings