Selection-free zinc-finger-nuclease engineering by context-dependent assembly (CoDA)

JD Sander, EJ Dahlborg, MJ Goodwin, L Cade… - Nature …, 2011 - nature.com
JD Sander, EJ Dahlborg, MJ Goodwin, L Cade, F Zhang, D Cifuentes, SJ Curtin
Nature methods, 2011nature.com
Engineered zinc-finger nucleases (ZFNs) enable targeted genome modification. Here we
describe context-dependent assembly (CoDA), a platform for engineering ZFNs using only
standard cloning techniques or custom DNA synthesis. Using CoDA-generated ZFNs, we
rapidly altered 20 genes in Danio rerio, Arabidopsis thaliana and Glycine max. The
simplicity and efficacy of CoDA will enable broad adoption of ZFN technology and make
possible large-scale projects focused on multigene pathways or genome-wide alterations.
Abstract
Engineered zinc-finger nucleases (ZFNs) enable targeted genome modification. Here we describe context-dependent assembly (CoDA), a platform for engineering ZFNs using only standard cloning techniques or custom DNA synthesis. Using CoDA-generated ZFNs, we rapidly altered 20 genes in Danio rerio, Arabidopsis thaliana and Glycine max. The simplicity and efficacy of CoDA will enable broad adoption of ZFN technology and make possible large-scale projects focused on multigene pathways or genome-wide alterations.
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