A rapid and general assay for monitoring endogenous gene modification.
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.
Guschin
Dmitry Y
DY
Sangamo BioSciences, Inc., Richmond, CA, USA.
Waite
Adam J
AJ
Katibah
George E
GE
Miller
Jeffrey C
JC
Holmes
Michael C
MC
Rebar
Edward J
EJ
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
D004720:Endonucleases