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

29 Citations (Scopus)

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

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Clustered Regularly Interspaced Short Palindromic Repeats
Gene expression
Guide RNA
Genes
Endonucleases
Gene Silencing
RNA
Fusion reactions
Networks (circuits)

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Yeo, N. C., Chavez, A., Lance-Byrne, A., Chan, Y., Menn, D., Milanova, D., ... 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

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

In: Nature Methods, Vol. 15, No. 8, 01.08.2018, p. 611-616.

Research output: Contribution to journalArticle

Yeo, NC, Chavez, A, Lance-Byrne, A, Chan, Y, Menn, D, Milanova, D, Kuo, CC, Guo, X, Sharma, S, Tung, A, Cecchi, RJ, Tuttle, M, Pradhan, S, Lim, ET, Davidsohn, N, Ebrahimkhani, MR, Collins, JJ, Lewis, NE, Kiani, S & Church, GM 2018, 'An enhanced CRISPR repressor for targeted mammalian gene regulation', Nature Methods, vol. 15, no. 8, pp. 611-616. https://doi.org/10.1038/s41592-018-0048-5
Yeo NC, Chavez A, Lance-Byrne A, Chan Y, Menn D, Milanova D et al. An enhanced CRISPR repressor for targeted mammalian gene regulation. Nature Methods. 2018 Aug 1;15(8):611-616. https://doi.org/10.1038/s41592-018-0048-5
Yeo, Nan Cher ; Chavez, Alejandro ; Lance-Byrne, Alissa ; Chan, Yingleong ; Menn, David ; Milanova, Denitsa ; Kuo, Chih Chung ; Guo, Xiaoge ; Sharma, Sumana ; Tung, Angela ; Cecchi, Ryan J. ; Tuttle, Marcelle ; Pradhan, Swechchha ; Lim, Elaine T. ; Davidsohn, Noah ; Ebrahimkhani, Mo R. ; Collins, James J. ; Lewis, Nathan E. ; Kiani, Samira ; Church, George M. / An enhanced CRISPR repressor for targeted mammalian gene regulation. In: Nature Methods. 2018 ; Vol. 15, No. 8. pp. 611-616.
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