Gene Correction pp 371-383

Part of the Methods in Molecular Biology book series (MIMB, volume 1114)

Identification of Off-Target Cleavage Sites of Zinc Finger Nucleases and TAL Effector Nucleases Using Predictive Models

  • Eli J. Fine
  • Thomas J. Cradick
  • Gang Bao
Protocol

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.

Key words

Gene targeting Off target effects Zinc finger nuclease ZFN Transcription activator-like effector nuclease TALEN Gene therapy Off-target sites Bioinformatics 

Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Eli J. Fine
    • 1
  • Thomas J. Cradick
    • 1
  • Gang Bao
    • 1
  1. 1.Wallace H. Coulter Department of Biomedical EngineeringGeorgia Institute of TechnologyAtlantaUSA

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