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

Nat Methods. 2011 Jan;8(1):67-9. doi: 10.1038/nmeth.1542. Epub 2010 Dec 12.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Arabidopsis / genetics
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Endonucleases / genetics*
  • Endonucleases / metabolism*
  • Genome
  • Protein Engineering*
  • Soybeans / genetics
  • Zebrafish / genetics
  • Zinc Fingers / genetics
  • Zinc Fingers / physiology*

Substances

  • DNA-Binding Proteins
  • Endonucleases