title: Small molecule-triggered Cas9 protein with improved genome-editing specificity.
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.
authors.affiliation: 1] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Davis
authors.affiliation: Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Vikram
authors.initials: V
authors.lastname: Pattanayak
authors.affiliation: 1] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: David B
authors.initials: DB
authors.lastname: Thompson
authors.affiliation: 1] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Zuris
authors.affiliation: 1] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA. [2] Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Liu
mesh_terms: D001426:Bacterial Proteins / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D004720:Endonucleases / Q000187:drug effects / Q000235:genetics*; D016678:Genome / Q000187:drug effects*; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D011487:Protein Conformation; D054852:Small Molecule Libraries; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D008024:Ligands; D054852:Small Molecule Libraries; D013629:Tamoxifen; C016601:afimoxifene; D004720:Endonucleases
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