Identification of off-target cleavage sites of zinc finger nucleases and TAL effector nucleases using predictive models

Methods Mol Biol. 2014:1114:371-83. doi: 10.1007/978-1-62703-761-7_24.

Abstract

Using engineered nucleases, such as Zinc Finger Nucleases (ZFNs) or Transcription Activator-Like Effector Nucleases (TALENs), to make targeted genomic modifications has become a common technique to create new model organisms and custom cell lines, and has shown great promise for disease treatment. However, these nucleases have the potential for off-target cleavage that could confound interpretation of experimental results and be detrimental for therapeutic use. Here, we describe a method to test for nuclease cleavage at potential off-target sites predicted by bioinformatics models.

Publication types

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

MeSH terms

  • Animals
  • Computational Biology / methods*
  • DNA Cleavage*
  • Endonucleases / metabolism*
  • Substrate Specificity
  • Zinc Fingers / physiology*

Substances

  • Endonucleases