Journal
NATURE METHODS
Volume 15, Issue 8, Pages 611-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41592-018-0048-5
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Funding
- NIH [RM1 HG008525, P50 HG005550]
- National Cancer Institute [5T32CA009216-34]
- Burroughs Wellcome Fund (Career Award for Medical Scientists)
- NIGMS [R35 GM119850]
- Novo Nordisk Foundation [NNF10CC1016517]
- DARPA (Young Faculty Award) [D16AP00047]
- Arizona State University Fulton Schools of Engineering startup fund
- Paul G. Allen Frontiers Group
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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.
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