Selection-free zinc-finger-nuclease engineering by context-dependent assembly (CoDA).
basic_science · Level V
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- Record sourced from PubMed, PMID 21151135.
- Also identified by DOI 10.1038/nmeth.1542 and PMC identifier 3018472.
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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.
Medical subject headings
- Endonucleases
- Protein Engineering
- Zinc Fingers