Off-target Effects in CRISPR/Cas9-mediated Genome Engineering.
Off-target Effects in CRISPR/Cas9-mediated Genome Engineering.
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DOI：
10.1038/mtna.2015.37
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发表时间：
2015-11-17
期刊：
影响因子：
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通讯作者：
Yang SH
中科院分区：
文献类型：
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作者：
Zhang XH;Tee LY;Wang XG;Huang QS;Yang SH
CRISPR/Cas9是一种通用的基因组编辑技术，广泛用于研究遗传元件的功能，创造转基因生物以及遗传疾病的临床前研究。然而，高频率的脱靶活性(≥50%)-RGEN （rna引导的核酸内切酶）-诱导突变发生在目标位点以外的其他位点是一个主要的问题，特别是在治疗和临床应用中。在这里，我们回顾了CRISPR/Cas9系统中脱靶切割的基本机制，检测脱靶突变的方法，以及最小化脱靶切割的策略。CRISPR/Cas9系统脱靶特异性的提高将提供坚实的基因型-表型相关性，从而实现对基因组编辑数据的忠实解释，这必将促进该技术的基础和临床应用。
CRISPR/Cas9 is a versatile genome-editing technology that is widely used for studying the functionality of genetic elements, creating genetically modified organisms as well as preclinical research of genetic disorders. However, the high frequency of off-target activity (≥50%)—RGEN (RNA-guided endonuclease)-induced mutations at sites other than the intended on-target site—is one major concern, especially for therapeutic and clinical applications. Here, we review the basic mechanisms underlying off-target cutting in the CRISPR/Cas9 system, methods for detecting off-target mutations, and strategies for minimizing off-target cleavage. The improvement off-target specificity in the CRISPR/Cas9 system will provide solid genotype–phenotype correlations, and thus enable faithful interpretation of genome-editing data, which will certainly facilitate the basic and clinical application of this technology.