title: A rapid and general assay for monitoring endogenous gene modification.
abstract: The development of zinc finger nucleases for targeted gene modification can benefit from rapid functional assays that directly quantify activity at the endogenous target. Here we describe a simple procedure for quantifying mutations that result from DNA double-strand break repair via non-homologous end joining. The assay is based on the ability of the Surveyor nuclease to selectively cleave distorted duplex DNA formed via cross-annealing of mutated and wild-type sequence.
authors.affiliation: Sangamo BioSciences, Inc., Richmond, CA, USA.
authors.forename: Dmitry Y
authors.initials: DY
authors.lastname: Guschin
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Waite
authors.forename: George E
authors.initials: GE
authors.lastname: Katibah
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Miller
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Rebar
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004591:Electrophoresis, Polyacrylamide Gel; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D016133:Polymerase Chain Reaction; D016335:Zinc Fingers / Q000235:genetics
chemical_list: D004720:Endonucleases