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

Nat Chem Biol

Authors	Kevin Davis Vikram Pattanayak David Thompson John Zuris David Liu
Keywords	Humans Genome HEK293 Cells Models, Molecular High-Throughput Nucleotide Sequencing Cells, Cultured Bacterial Proteins Endonucleases Ligands Protein Conformation Small Molecule Libraries Tamoxifen
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.
Year of Publication	2015
Journal	Nat Chem Biol
Volume	11
Issue	5
Pages	316-8
Date Published	2015 May
ISSN	1552-4469
DOI	10.1038/nchembio.1793
PubMed ID	25848930
PubMed Central ID	PMC4402137
Links
Grant list	F32 GM 106601-2 / GM / NIGMS NIH HHS / United States T32 GM007753 / GM / NIGMS NIH HHS / United States Howard Hughes Medical Institute / United States F32 GM106601 / GM / NIGMS NIH HHS / United States R01 GM095501 / GM / NIGMS NIH HHS / United States