A rapid and general assay for monitoring endogenous gene modification
Engineered Zinc Finger Proteins: Methods and Protocols, 2010•Springer
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.
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.
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.
Springer