Ma Xing-liang, Liu Yao-guang
1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangzhou 510642, China; 2. College of Life Sciences, South China Agricultural University, Guangzhou 510642, China;
Yi Chuan. 2016 Feb;38(2):118-25. doi: 10.16288/j.yczz.15-395.
Targeted genomic editing technologies use programmable DNA nucleases to cleave genomic target sites, thus inducing targeted mutations in the genomes. The newly prevailed clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system that consists of the Cas9 nuclease and single guide RNA (sgRNA) has the advantages of simplicity and high efficiency as compared to other programmable DNA nuclease systems such as zinc finger nucleases (ZFNs) and transcription activator like effector nucleases (TALENs). Currently, a number of cases have been reported on the application of the CRISPR/Cas9 genomic editing technology in plants. In this review, we summarize the strategies for preparing the Cas9 and sgRNA expression constructs, the transformation method for obtaining targeted mutations, the efficiency and features of the resulting mutations and the methods for detecting or genotyping of the mutation sites. We also discuss the existing problems and perspectives of CRISPR/Cas9-based genomic editing in plants.
靶向基因组编辑技术利用可编程的DNA核酸酶切割基因组靶位点，从而在基因组中诱导靶向突变。与其他可编程DNA核酸酶系统（如锌指核酸酶（ZFNs）和转录激活因子样效应物核酸酶（TALENs））相比，新流行的由Cas9核酸酶和单向导RNA（sgRNA）组成的成簇规律间隔短回文重复序列/CRISPR相关蛋白9（CRISPR/Cas9）系统具有简单高效的优点。目前，已有许多关于CRISPR/Cas9基因组编辑技术在植物中应用的报道。在本综述中，我们总结了制备Cas9和sgRNA表达构建体的策略、获得靶向突变的转化方法、所得突变的效率和特征以及突变位点的检测或基因分型方法。我们还讨论了基于CRISPR/Cas9的植物基因组编辑中存在的问题和前景。
