Minimizing off-Target Mutagenesis Risks Caused by Programmable Nucleases.
Minimizing off-Target Mutagenesis Risks Caused by Programmable Nucleases.
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DOI：
10.3390/ijms161024751
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发表时间：
2015-10-16
影响因子：
5.6
通讯作者：
Hotta A
中科院分区：
文献类型：
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作者：
Ishida K;Gee P;Hotta A
可编程核酸酶，如锌指核酸酶（ZFNs）、转录激活因子样效应核酸酶（TALENs）和聚集规律穿插的短回语重复相关蛋白-9 (CRISPR-Cas9)，在治疗遗传疾病或预防传染病的临床应用中具有巨大的潜力。然而，由于这些核酸酶对DNA识别的准确性并不总是完美的，脱靶突变可能导致治疗患者出现不良事件，如细胞毒性或肿瘤发生。因此，设计核酸酶和分析其活性必须仔细评估，以尽量减少脱靶突变。此外，严格的基因组检测对于确保核酸酶修饰细胞的完整性至关重要。在这篇综述中，我们概述了现有的核酸酶设计平台、核酸酶工程方法以最小化脱靶活性，以及评估CRISPR-Cas9的靶向和脱靶切割的方法。
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.