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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.

Original publication

DOI

10.1038/s41592-018-0048-5

Type

Journal article

Journal

Nature methods

Publication Date

08/2018

Volume

15

Pages

611 - 616

Addresses

Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA, USA.

Keywords

Humans, Recombinant Fusion Proteins, Repressor Proteins, RNA, Guide, Gene Expression Regulation, Gene Silencing, Genes, Synthetic, Methyl-CpG-Binding Protein 2, HEK293 Cells, CRISPR-Cas Systems, CRISPR-Associated Protein 9