Small molecule-triggered Cas9 protein with improved genome-editing specificity

Nat Chem Biol. 2015 May;11(5):316-8. doi: 10.1038/nchembio.1793. Epub 2015 Apr 6.

Abstract

Directly modulating the activity of genome-editing proteins has the potential to increase their specificity by reducing activity following target locus modification. We developed Cas9 nucleases that are activated by the presence of a cell-permeable small molecule by inserting an evolved 4-hydroxytamoxifen-responsive intein at specific positions in Cas9. In human cells, conditionally active Cas9s modify target genomic sites with up to 25-fold higher specificity than wild-type Cas9.

Publication types

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

MeSH terms

  • Bacterial Proteins / drug effects
  • Bacterial Proteins / genetics
  • Cells, Cultured
  • Endonucleases / drug effects
  • Endonucleases / genetics*
  • Genome / drug effects*
  • HEK293 Cells
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Ligands
  • Models, Molecular
  • Protein Conformation
  • Small Molecule Libraries
  • Tamoxifen / analogs & derivatives
  • Tamoxifen / pharmacology

Substances

  • Bacterial Proteins
  • Ligands
  • Small Molecule Libraries
  • Tamoxifen
  • afimoxifene
  • Endonucleases