title: Continuous directed evolution of DNA-binding proteins to improve TALEN specificity.
abstract: Nucleases containing programmable DNA-binding domains can alter the genomes of model organisms and have the potential to become human therapeutics. Here we present DNA-binding phage-assisted continuous evolution (DB-PACE) as a general approach for the laboratory evolution of DNA-binding activity and specificity. We used this system to generate transcription activator-like effectors nucleases (TALENs) with broadly improved DNA cleavage specificity, establishing DB-PACE as a versatile approach for improving the accuracy of genome-editing agents.
authors.affiliation: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: Basil P
authors.initials: BP
authors.lastname: Hubbard
authors.affiliation: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: Ahmed H
authors.identifier: http://orcid.org/0000-0002-8105-1883
authors.initials: AH
authors.lastname: Badran
authors.affiliation: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Zuris
authors.affiliation: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Guilinger
authors.affiliation: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Davis
authors.affiliation: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: Liwei
authors.initials: L
authors.lastname: Chen
authors.affiliation: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Center for Cancer Research, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Shengdar Q
authors.initials: SQ
authors.lastname: Tsai
authors.affiliation: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Center for Cancer Research, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Jeffry D
authors.initials: JD
authors.lastname: Sander
authors.affiliation: Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Center for Cancer Research, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: J Keith
authors.initials: JK
authors.lastname: Joung
authors.affiliation: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.|Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Liu
mesh_terms: D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism*; D019020:Directed Molecular Evolution / Q000379:methods*; D018390:Gene Targeting / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D063150:Polycomb Repressive Complex 1 / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods
chemical_list: C570428:CBX8 protein, human; D004268:DNA-Binding Proteins; D063150:Polycomb Repressive Complex 1; D003851:Deoxyribonucleases