工程化引导 RNA 以减少 CRISPR 的脱靶效应。
Engineering guide RNA to reduce the off-target effects of CRISPR.
机构信息
Department of Pathology, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China; Frontier Science Center for Immunology and Metabolism, Wuhan University, 430071, China.
Department of Pathology, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China; Frontier Science Center for Immunology and Metabolism, Wuhan University, 430071, China; Department of Urology, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China.
出版信息
J Genet Genomics. 2019 Nov 20;46(11):523-529. doi: 10.1016/j.jgg.2019.11.003. Epub 2019 Nov 23.
As versatile and robust genome editing tools, clustered regularly interspaced short palindromic repeats (CRISPR) technologies have been broadly used in basic research, biotechnology, and therapeutic development. Off-target mutagenesis by CRISPR systems has been demonstrated, and various methods have been developed to markedly increase their specificity. In this review, we highlight the efforts of producing and modifying guide RNA (gRNA) to minimize off-target activities, including sequence and structure design, tuning expression and chemical modification. The modalities of gRNA engineering can be applied across CRISPR systems. In conjunction with CRISPR protein effectors, the engineered gRNA enables efficient and precise genome editing.
作为多功能且强大的基因组编辑工具，成簇规律间隔短回文重复序列（CRISPR）技术已广泛应用于基础研究、生物技术和治疗开发。CRISPR 系统的脱靶突变已被证明，并且已经开发了各种方法来显著提高其特异性。在这篇综述中，我们强调了产生和修饰指导 RNA（gRNA）以最小化脱靶活性的努力，包括序列和结构设计、表达调控和化学修饰。gRNA 工程的方式可以应用于各种 CRISPR 系统。与 CRISPR 蛋白效应物结合，工程化的 gRNA 可实现高效、精确的基因组编辑。
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