Efficiency Optimization of CRISPR/Cas9-Mediated Targeted Mutagenesis in Grape
文献类型：期刊论文
| 作者 | Ren, Fengrui1,3; Ren, Chong1,3; Zhang, Zhan1,3; Duan, Wei1; Lecourieux, David4; Li, Shaohua1,3; Liang, Zhenchang2,3 |
| 刊名 | FRONTIERS IN PLANT SCIENCE
 |
| 出版日期 | 2019-05-16 |
| 卷号 | 10页码:9 |
| 关键词 | CRISPR/Cas9 optimization GC content grape gene expression gene editing efficiency |
| ISSN号 | 1664-462X |
| DOI | 10.3389/fpls.2019.00612 |
| 英文摘要 | Clustered regularly interspersed short palindromic repeats (CRISPR)/Cas system is an efficient targeted genome editing method. Although CRISPR/Cas9-mediated mutagenesis has been applied successfully in grape, few studies have examined the technique's efficiency. To optimize CRISPR/Cas9 editing efficiency in Vitis vinifera, we surveyed three key parameters: GC content of single guide RNA (sgRNA), variety of transformant cells used, and SpCas9 expression levels in transgenic cell mass. Four sgRNAs with differing GC content were designed to target exon sites of the V. vinifera phytoene desaturase gene. Suspension cells of 'Chardonnay' and '41B' varieties were used as the transgenic cell mass. Both T7EI and PCR/RE assays showed that CRISPR/Cas9 editing efficiency increases proportionally with sgRNA GC content with 65% GC content yielding highest editing efficiency in both varieties. Additionally, gene editing was more efficient in '41B' than in 'Chardonnay.' CRISPR/Cas9 systems with different editing efficiency showed different SpCas9 expression level, but compared with GC content of sgRNA, SpCas9 expression level has less influence on editing efficiency. Taken together, these results help optimize of CRISPR/Cas9 performance in grape. |
| 资助项目 | National Key Research and Development Program of China ; STS project of Chinese Academy of Sciences[KFJ-STS-ZDTP-025] ; Major Science and Technology Program of Ningxia Hui Autonomous Region[2016BZ06] ; Agricultural Breeding Project of Ningxia Hui Autonomous Region[NXNYYZ20150203] ; National Natural Science Foundation of China[31772266] |
| WOS研究方向 | Plant Sciences |
| 语种 | 英语 |
| WOS记录号 | WOS:000468131600001 |
| 出版者 | FRONTIERS MEDIA SA |
| 源URL | [http://202.127.146.157/handle/2RYDP1HH/7726]  |
| 专题 | 中国科学院武汉植物园 |
| 通讯作者 | Liang, Zhenchang |
| 作者单位 | 1.Univ Chinese Acad Sci, Coll Life Sci, Beijing, Peoples R China 2.Chinese Acad Sci, Sino Africa Joint Res Ctr, Wuhan, Hubei, Peoples R China 3.Chinese Acad Sci, Inst Bot, Lab Plant Resources, Beijing Key Lab Grape Sci & Enol, Beijing, Peoples R China 4.Univ Bordeaux, INRA, Bordeaux Sci Agro, EGFV, Villenave Dornon, France |
| 推荐引用方式 GB/T 7714 | Ren, Fengrui,Ren, Chong,Zhang, Zhan,et al. Efficiency Optimization of CRISPR/Cas9-Mediated Targeted Mutagenesis in Grape[J]. FRONTIERS IN PLANT SCIENCE,2019,10:9. |
| APA | Ren, Fengrui.,Ren, Chong.,Zhang, Zhan.,Duan, Wei.,Lecourieux, David.,...&Liang, Zhenchang.(2019).Efficiency Optimization of CRISPR/Cas9-Mediated Targeted Mutagenesis in Grape.FRONTIERS IN PLANT SCIENCE,10,9. |
| MLA | Ren, Fengrui,et al."Efficiency Optimization of CRISPR/Cas9-Mediated Targeted Mutagenesis in Grape".FRONTIERS IN PLANT SCIENCE 10(2019):9. |
入库方式： OAI收割
来源：武汉植物园
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