A Rapid and General Assay for Monitoring Endogenous Gene Modification

0301 basic medicine 0303 health sciences 03 medical and health sciences DNA Repair Animals Humans Biological Assay DNA Breaks, Double-Stranded Electrophoresis, Polyacrylamide Gel Zinc Fingers Endonucleases Models, Biological Polymerase Chain Reaction
DOI: 10.1007/978-1-60761-753-2_15 Publication Date: 2010-08-02T20:14:58Z
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.
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