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
AUTHORS (6)
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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