Minimizing off-Target Mutagenesis Risks Caused by Programmable Nucleases

International Journal of Molecular Sciences

Kentaro Ishida¹  Peter Gee¹  Akitsu Hotta¹ 

[1]Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan

[2] E-Mails:

关键词: CRISPR Cas9;    genome editing;    mutagenesis;    off-target effect;   

DOI  :  10.3390/ijms161024751

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.