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Minimizing off-Target Mutagenesis Risks Caused by Programmable Nucleases

Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan
Institute for Integrated Cell Material Sciences (iCeMS), Kyoto University, Yoshida Ushinomiya-cho, Sakyo-ku, Kyoto 606-8507, Japan
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Izuho Hatada
Int. J. Mol. Sci. 2015, 16(10), 24751-24771;
Received: 26 August 2015 / Revised: 2 October 2015 / Accepted: 8 October 2015 / Published: 16 October 2015
(This article belongs to the Special Issue Genome Editing)
Programmable nucleases, such as zinc finger nucleases (ZFNs), transcription activator like effector nucleases (TALENs), and clustered regularly interspersed short palindromic repeats associated protein-9 (CRISPR-Cas9), hold tremendous potential for applications in the clinical setting to treat genetic diseases or prevent infectious diseases. However, because the accuracy of DNA recognition by these nucleases is not always perfect, off-target mutagenesis may result in undesirable adverse events in treated patients such as cellular toxicity or tumorigenesis. Therefore, designing nucleases and analyzing their activity must be carefully evaluated to minimize off-target mutagenesis. Furthermore, rigorous genomic testing will be important to ensure the integrity of nuclease modified cells. In this review, we provide an overview of available nuclease designing platforms, nuclease engineering approaches to minimize off-target activity, and methods to evaluate both on- and off-target cleavage of CRISPR-Cas9. View Full-Text
Keywords: CRISPR Cas9; genome editing; mutagenesis; off-target effect CRISPR Cas9; genome editing; mutagenesis; off-target effect
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Ishida, K.; Gee, P.; Hotta, A. Minimizing off-Target Mutagenesis Risks Caused by Programmable Nucleases. Int. J. Mol. Sci. 2015, 16, 24751-24771.

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