植物CRISPR/Cas9基因组编辑系统与突变分析

doi:10.16288/j.yczz.15-395

遗传 ›› 2016, Vol. 38 ›› Issue (2): 118-125.doi: 10.16288/j.yczz.15-395

植物CRISPR/Cas9基因组编辑系统与突变分析

马兴亮^1,2,刘耀光^1,2

1. 亚热带农业生物资源保护与利用国家重点实验室,广州 510642;
2. 华南农业大学生命科学学院,广州 510642

收稿日期:2015-09-15 修回日期:2015-12-03 出版日期:2016-02-20 发布日期:2015-12-30
通讯作者: 刘耀光,博士,研究员,博士生导师,研究方向：植物遗传学与分子生物学。E-mail: ygliu@scau.edu.cn
作者简介:马兴亮,博士研究生,研究方向：生物化学与分子生物学。E-mail: xlma@stu.scau.edu.cn
基金资助:
转基因植物研究与产业化专项(编号：2014ZX08010-001, 2014ZX08009-002)资助[Supported by the Transgenic Research Program (Nos.2014ZX08010-001, 2014ZX08009-002); Ministry of Agriculture of the People’s Republic of China]

CRISPR/Cas9-based genome editing systems and the analysis of targeted genome mutations in plants

Xingliang Ma^{1, 2}, Yaoguang Liu^{1, 2}

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

Received:2015-09-15 Revised:2015-12-03 Online:2016-02-20 Published:2015-12-30

摘要/Abstract

摘要： 基因组定点编辑技术通过可编码核酸酶切割基因组特定位点,进而诱导基因组定点突变。CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9)基因组编辑系统由Cas9核酸酶以及sgRNA(Single guide RNA)组成,与其他可编码核酸酶系统如锌指核酸酶(Zinc finger nucleases, ZFNs)和类转录激活因子效应物核酸酶(Transcription activator-like effector nucleases, TALENs)相比具有更简便的操作性和更高的基因组定点编辑效率。目前在植物中已有多例应用CRISPR/Cas9系统进行基因组编辑的报道。本文从Cas9基因与sgRNA表达载体的构建策略,获得基因组定点编辑突变体的转化方法、突变的效率和特征、突变的检测方法等方面进行了总结,最后对植物中利用CRISPR/Cas9系统进行基因组编辑存在的问题以及发展前景进行了讨论。

关键词: 植物, CRISPR/Cas9, 靶向突变, 基因型鉴定

Abstract: 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.

Key words: plants, CRISPR/Cas9, targeted mutations, genotyping

马兴亮,刘耀光. 植物CRISPR/Cas9基因组编辑系统与突变分析[J]. 遗传, 2016, 38(2): 118-125.

Xingliang Ma, Yaoguang Liu. CRISPR/Cas9-based genome editing systems and the analysis of targeted genome mutations in plants[J]. HEREDITAS(Beijing), 2016, 38(2): 118-125.

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植物CRISPR/Cas9基因组编辑系统与突变分析

CRISPR/Cas9-based genome editing systems and the analysis of targeted genome mutations in plants

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