Repositorium
Systematic identification of endogenous RNA polymerase III promoters for efficient RNA guide-based genome editing technologies in maize
337
Journal Article / 2018
Qi, Xiantao; Le Dong; Liu, Changlin; Mao, Long; Liu, Fang; Zhang, Xin; Cheng, Beijiu; Xie, Chuanxiao
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 Based on the reported and unpublished data, the in vitro TLR assay results were confirmed to be a readily system and may be extended to other plant species amenable to efficient genome editing via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
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
793 |
Xie, Chuanxiao China |
Zea mays |
CRISPR/Cas9 Wx1 |
Waxy phenotype SDN1 |
Market-oriented Product quality |