Engineering designer nucleases with customized cleavage specificities

Curr Protoc Mol Biol. 2011 Oct:Chapter 12:Unit12.13. doi: 10.1002/0471142727.mb1213s96.

Abstract

Engineered designer nucleases can be used to efficiently modify genomic sequence in a wide variety of model organisms and cell types. Zinc finger nucleases (ZFNs), consisting of an engineered zinc finger array fused to a non-specific cleavage domain, have been extensively used to modify a broad range of endogenous genes. Protocols for engineering ZFNs targeted to specific gene sequences of interest using the context-dependent assembly (CoDA) method are described in this unit.

MeSH terms

  • DNA / metabolism*
  • Deoxyribonucleases / genetics*
  • Deoxyribonucleases / metabolism*
  • Genetic Engineering / methods*
  • Protein Binding
  • Substrate Specificity
  • Zinc Fingers / genetics

Substances

  • DNA
  • Deoxyribonucleases