Rani Reema, Yadav Prashant, Barbadikar Kalyani M, Baliyan Nikita, Malhotra Era Vaidya, Singh Binay Kumar, Kumar Arun, Singh Dhiraj
ICAR-Directorate of Rapeseed-Mustard Research, Bharatpur, Rajasthan, 321303, India.
ICAR-Indian Institute of Rice Research, Hyderabad, Telangana, India.
Biotechnol Lett. 2016 Dec;38(12):1991-2006. doi: 10.1007/s10529-016-2195-z. Epub 2016 Aug 29.
Creation of variation in existing gene pool of crop plants is the foremost requirement in crop improvement programmes. Genome editing is a tool to produce knock out of target genes either by introduction of insertion or by deletion that disrupts the function of a specific gene. The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) system is the most recent addition to the toolbox of sequence-specific nucleases that includes ZFNs and TALENs. The CRISPR/Cas9 system allows targeted cleavage of genomic DNA guided by a small noncoding RNA, resulting in gene modifications by both non-homologous end joining and homology-directed repair mechanisms. Here, we present an overview of mechanisms of CRISPR, its potential roles in creating variation in germplasm and applications of this novel interference pathway in crop improvement. The availability of the CRISPR/Cas9 system holds promise in facilitating both forward and reverse genetics and will enhance research in crops that lack genetic resources.
在作物改良计划中,首要需求是在现有作物植物基因库中创造变异。基因组编辑是一种通过引入插入或缺失来产生目标基因敲除的工具,这些插入或缺失会破坏特定基因的功能。CRISPR/Cas9(成簇规律间隔短回文重复序列/CRISPR相关蛋白9)系统是序列特异性核酸酶工具库中的最新成员,该工具库还包括锌指核酸酶(ZFNs)和转录激活样效应因子核酸酶(TALENs)。CRISPR/Cas9系统允许在一个小的非编码RNA引导下对基因组DNA进行靶向切割,通过非同源末端连接和同源定向修复机制实现基因修饰。在此,我们概述了CRISPR的机制、其在种质变异创造中的潜在作用以及这种新型干扰途径在作物改良中的应用。CRISPR/Cas9系统的可用性有望促进正向和反向遗传学研究,并将加强对缺乏遗传资源作物的研究。