Kharbikar Lalit, Konwarh Rocktotpal, Chakraborty Monoswi, Nandanwar Shweta, Marathe Ashish, Yele Yogesh, Ghosh Probir Kumar, Sanan-Mishra Neeti, Singh Anand Pratap
ICAR - National Institute of Biotic Stress Management (NIBSM), Raipur, India.
International Centre for Genetic Engineering and Biotechnology (ICGEB), New Delhi, India.
Physiol Mol Biol Plants. 2023 Dec;29(12):1825-1850. doi: 10.1007/s12298-023-01397-3. Epub 2023 Dec 7.
The recent thrust in research has projected the type II clustered regularly interspaced short palindromic repeats and associated protein 9 (CRISPR-Cas9) system as an avant-garde plant genome editing tool. It facilitates the induction of site-specific double-stranded DNA cleavage by the RNA-guided DNA endonuclease (RGEN), Cas9. Elimination, addition, or alteration of sections in DNA sequence besides the creation of a knockout genotype (CRISPRko) is aided by the CRISPR-Cas9 system in its wild form (wtCas9). The inactivation of the nuclease domain generates a dead Cas9 (dCas9), which is capable of targeting genomic DNA without scissoring it. The dCas9 system can be engineered by fusing it with different effectors to facilitate transcriptional activation (CRISPRa) and transcriptional interference (CRISPRi). CRISPR-Cas thus holds tremendous prospects as a genome-manipulating stratagem for a wide gamut of crops. In this article, we present a brief on the fundamentals and the general workflow of the CRISPR-Cas system followed by an overview of the prospects of bioinformatics in propelling CRISPR-Cas research with a special thrust on the available databases and algorithms/web-accessible applications that have aided in increasing the usage and efficiency of editing. The article also provides an update on the current regulatory landscape in different countries on the CRISPR-Cas edited plants to emphasize the far-reaching impact of the genomic editing technology.
The online version contains supplementary material available at 10.1007/s12298-023-01397-3.
近期的研究重点已将II型成簇规律间隔短回文重复序列及其相关蛋白9(CRISPR-Cas9)系统作为一种前沿的植物基因组编辑工具。它通过RNA引导的DNA内切酶(RGEN)Cas9促进位点特异性双链DNA切割的诱导。除了创建敲除基因型(CRISPRko)外,CRISPR-Cas9系统的野生型(wtCas9)有助于消除、添加或改变DNA序列中的片段。核酸酶结构域的失活产生了一种无核酸酶活性的Cas9(dCas9),它能够靶向基因组DNA而不进行切割。dCas9系统可以通过与不同的效应器融合进行改造,以促进转录激活(CRISPRa)和转录干扰(CRISPRi)。因此,CRISPR-Cas作为一种用于多种作物的基因组操作策略具有巨大的前景。在本文中,我们简要介绍了CRISPR-Cas系统的基本原理和一般工作流程,然后概述了生物信息学在推动CRISPR-Cas研究方面的前景,特别强调了有助于提高编辑使用和效率的可用数据库以及算法/网络可访问应用程序。本文还提供了不同国家对CRISPR-Cas编辑植物的当前监管情况的更新,以强调基因组编辑技术的深远影响。
在线版本包含可在10.1007/s12298-023-01397-3获取的补充材料。