An enhanced CRISPR repressor for targeted mammalian gene regulation

Nat Methods. 2018 Aug;15(8):611-616. doi: 10.1038/s41592-018-0048-5. Epub 2018 Jul 16.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

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


  • MECP2 protein, human
  • Methyl-CpG-Binding Protein 2
  • RNA, Guide, CRISPR-Cas Systems
  • Recombinant Fusion Proteins
  • Repressor Proteins
  • CRISPR-Associated Protein 9