清脆的
基因组编辑
突变体
解码方法
基因
协议(科学)
遗传学
计算生物学
Cas9
生物
序列(生物学)
计算机科学
医学
算法
病理
替代医学
作者
Dongchang Zeng,Xingliang Ma,Xianrong Xie,Qinlong Zhu,Yao-Guang Liu
出处
期刊:aZhongguo kexue
[Science in China Press]
日期:2018-06-21
卷期号:48 (7): 783-794
被引量:11
标识
DOI:10.1360/n052018-00069
摘要
Genome editing with CRISPR/Cas9 has become an effective tool for gene functionality dissection and crop improvement. Accordingly, we have developed a CRISPR/Cas9 vector system for robust multi-gene editing in plants. Our inventory includes six versatile binary vectors and 12 intermediary sgRNA vectors with different U3 and U6 promoters, to achieve high efficient multi-gene editing, under specific antibiotic selections in both monocot and dicot plants. Additionally, we also have developed an integrated web-based software toolkit, CRISPR-GE, to facilitate the utilization CRISPR/Cas9 in plants. In this protocol, we provide detailed procedures for using the CRISPR/Cas9 system, including target site selection, construction of CRISPR/Cas9 binary vectors, Sanger-sequencing-based genotyping of mutant sites, and troubleshooting for common problems.
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