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.
Journal article
2018-08-01T00:00:00+00:00
15
611 - 616
5
CRISPR-Associated Protein 9, CRISPR-Cas Systems, Gene Expression Regulation, Gene Silencing, Genes, Synthetic, HEK293 Cells, Humans, Methyl-CpG-Binding Protein 2, RNA, Guide, CRISPR-Cas Systems, Recombinant Fusion Proteins, Repressor Proteins