An enhanced CRISPR repressor for targeted mammalian gene regulation

Nature Methods
Nan Cher YeoGeorge M Church

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.

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Citations

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Methods Mentioned

BETA
RNA-seq
flow cytometry
flow
transfection
PCR
transfections

Software Mentioned

limma
FlowJo
HtSeq
custom R scripts
EdgeR
Cutadapt
MAGeCK
STAR
TapeStation

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