Engineered anti-CRISPR proteins for optogenetic control of CRISPR-Cas9

Nat Methods. 2018 Nov;15(11):924-927. doi: 10.1038/s41592-018-0178-9. Epub 2018 Oct 30.

Abstract

Anti-CRISPR proteins are powerful tools for CRISPR-Cas9 regulation; the ability to precisely modulate their activity could facilitate spatiotemporally confined genome perturbations and uncover fundamental aspects of CRISPR biology. We engineered optogenetic anti-CRISPR variants comprising hybrids of AcrIIA4, a potent Streptococcus pyogenes Cas9 inhibitor, and the LOV2 photosensor from Avena sativa. Coexpression of these proteins with CRISPR-Cas9 effectors enabled light-mediated genome and epigenome editing, and revealed rapid Cas9 genome targeting in human cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques*
  • CRISPR-Associated Proteins / antagonists & inhibitors*
  • CRISPR-Cas Systems*
  • Epigenomics
  • Gene Editing*
  • Genome
  • HEK293 Cells
  • Humans
  • Light
  • Optogenetics*
  • Phototropins / chemistry*
  • Protein Engineering*
  • Streptococcus pyogenes / enzymology

Substances

  • CRISPR-Associated Proteins
  • Phototropins