An enhanced CRISPR repressor for targeted mammalian gene regulation

Nan Cher Yeo, Alejandro Chavez, Alissa Lance-Byrne, Yingleong Chan, David Menn, Denitsa Milanova, Chih Chung Kuo, Xiaoge Guo, Sumana Sharma, Angela Tung, Ryan J. Cecchi, Marcelle Tuttle, Swechchha Pradhan, Elaine T. Lim, Noah Davidsohn, Mo R. Ebrahimkhani, James J. Collins, Nathan E. Lewis, Samira Kiani, George M. Church

Research output: Contribution to journalArticle

43 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)611-616
Number of pages6
JournalNature Methods
Volume15
Issue number8
DOIs
StatePublished - Aug 1 2018

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Yeo, N. C., Chavez, A., Lance-Byrne, A., Chan, Y., Menn, D., Milanova, D., Kuo, C. C., Guo, X., Sharma, S., Tung, A., Cecchi, R. J., Tuttle, M., Pradhan, S., Lim, E. T., Davidsohn, N., Ebrahimkhani, M. R., Collins, J. J., Lewis, N. E., Kiani, S., & Church, G. M. (2018). An enhanced CRISPR repressor for targeted mammalian gene regulation. Nature Methods, 15(8), 611-616. https://doi.org/10.1038/s41592-018-0048-5