生物
清脆的
亚基因组mRNA
基因组编辑
发起人
Cas9
引导RNA
核糖核酸
遗传学
基因组
RNA聚合酶Ⅲ
基因
计算生物学
抄写(语言学)
RNA聚合酶
基因表达
哲学
语言学
作者
Xiantao Qi,Le Dong,Changlin Liu,Long Mao,Fang Liu,Xin Zhang,Beijiu Cheng,Chuanxiao Xie
出处
期刊:Crop Journal
[Elsevier]
日期:2018-06-01
卷期号:6 (3): 314-320
被引量:25
标识
DOI:10.1016/j.cj.2018.02.005
摘要
Single-guide RNA (sgRNA) is one of the two core components of the CRISPR (clustered regularly interspaced short palindromic repeat)/Cas (CRISPR-associated) genome-editing technology. We established an in vitro Traffic Light Reporter (TLR) system, which is designated as the same colors as traffic lights such as green, red and yellow were produced in cells. The TLR can be readily used in maize mesophyll protoplast for a quick test of promoter activity. The TLR assay indicates the variation in transcription activities of the seven Pol III promoters, from 3.4% (U6-1) to over 21.0% (U6-6). The U6-2 promoter, which was constructed to drive sgRNA expression targeting the ZmWx1 gene, yielded mutation efficiencies ranging from 48.5% to 97.1%. Based on the reported and unpublished data, the in vitro TLR assay results were confirmed to be a readily system and may be extended to other plant species amenable to efficient genome editing via CRISPR/Cas. Our efforts provide an efficient method of identifying native Pol III-recognized promoters for RNA guide-based genome-editing systems in maize.
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