Infectious Diseases and Immunology Division, CSIR-Indian Institute of Chemical Biology, Kolkatta, India.
Department of Botany, Visva-Bharati, West Bengal, India.
Prog Mol Biol Transl Sci. 2021;178:85-98. doi: 10.1016/bs.pmbts.2020.12.001. Epub 2021 Jan 21.
The discovery of CRISPR-Cas9 system has revolutionized the genome engineering research and has been established as a gold standard genome editing platform. This system has found its application in biochemical researches as well as in medical fields including disease diagnosis, development of therapeutics, etc. The enormous versatility of the CRISPR-Cas9 as a high throughput genome engineering platform, is derailed by its off-target activity. To overcome this, researchers from all over the globe have explored the system structurally and functionally and postulated several strategies to upgrade the system components including redesigning of Cas9 Nuclease and modification of guide RNA(gRNA) structure and customization of the protospacer adjacent motif. Here in this review, we portray the comprehensive overview of the strategies that has been adopted for redesigning the CRISPR-Cas9 system to enhance the efficiency and fidelity of the technology.
CRISPR-Cas9 系统的发现彻底改变了基因组工程研究，已成为一种标准的基因组编辑平台。该系统已应用于生化研究以及医学领域，包括疾病诊断、治疗药物的开发等。CRISPR-Cas9 作为高通量基因组工程平台具有巨大的多功能性，但也存在脱靶活性的问题。为了解决这个问题，来自世界各地的研究人员从结构和功能上对该系统进行了探索，并提出了几种策略来升级系统组件，包括 Cas9 核酸酶的重新设计、引导 RNA(gRNA)结构的修饰以及原间隔序列邻近基序的定制。在这篇综述中，我们全面概述了为改进 CRISPR-Cas9 系统以提高该技术的效率和准确性而采用的策略。
