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Measuring and Reducing Off-Target Activities of Programmable Nucleases Including CRISPR-Cas9

  • Koo, Taeyoung (Center for Genome Engineering, Institute for Basic Science) ;
  • Lee, Jungjoon (The Institute of Molecular Biology and Genetics) ;
  • Kim, Jin-Soo (Center for Genome Engineering, Institute for Basic Science)
  • Received : 2015.04.17
  • Accepted : 2015.04.30
  • Published : 2015.06.30

Abstract

Programmable nucleases, which include zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and RNA-guided engineered nucleases (RGENs) repurposed from the type II clustered, regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) system are now widely used for genome editing in higher eukaryotic cells and whole organisms, revolutionising almost every discipline in biological research, medicine, and biotechnology. All of these nucleases, however, induce off-target mutations at sites homologous in sequence with on-target sites, limiting their utility in many applications including gene or cell therapy. In this review, we compare methods for detecting nuclease off-target mutations. We also review methods for profiling genome-wide off-target effects and discuss how to reduce or avoid off-target mutations.

Acknowledgement

Supported by : IBS

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