Knowledge Management System Of Institute Of Botany,CAS
Efficiency Optimization of CRISPR/Cas9-Mediated Targeted Mutagenesis in Grape | |
Ren, Fengrui1; Ren, Chong1; Zhang, Zhan1; Duan, Wei1; Lecourieux, David2; Li, Shaohua1; Liang, Zhenchang3 | |
2019 | |
发表期刊 | FRONTIERS IN PLANT SCIENCE |
ISSN | 1664-462X |
卷号 | 10 |
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. |
关键词 | CRISPR/Cas9 optimization GC content grape gene expression gene editing efficiency |
学科领域 | Plant Sciences |
DOI | 10.3389/fpls.2019.00612 |
收录类别 | SCI |
语种 | 英语 |
WOS关键词 | GENETIC-TRANSFORMATION ; GENOME ; PLANTS ; ARABIDOPSIS ; GENERATION ; MULTIPLEX ; BINDING |
WOS研究方向 | Plant Sciences |
WOS记录号 | WOS:000468131600001 |
出版者 | FRONTIERS MEDIA SA |
文献子类 | Article |
出版地 | LAUSANNE |
资助机构 | 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 ChinaNational Natural Science Foundation of China (NSFC) [31772266] |
作者邮箱 | ZL249@ibcas.ac.cn |
作品OA属性 | gold, Green Published |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://ir.ibcas.ac.cn/handle/2S10CLM1/19605 |
专题 | 中科院北方资源植物重点实验室 |
作者单位 | 1.Chinese Acad Sci, Inst Bot, Lab Plant Resources, Beijing Key Lab Grape Sci & Enol, Beijing, Peoples R China 2.Univ Chinese Acad Sci, Coll Life Sci, Beijing, Peoples R China 3.Univ Bordeaux, INRA, Bordeaux Sci Agro, EGFV, Villenave Dornon, France 4.Chinese Acad Sci, Sino Africa Joint Res Ctr, Wuhan, Hubei, Peoples R China |
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. |
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. |
MLA | Ren, Fengrui,et al."Efficiency Optimization of CRISPR/Cas9-Mediated Targeted Mutagenesis in Grape".FRONTIERS IN PLANT SCIENCE 10(2019). |
条目包含的文件 | ||||||
文件名称/大小 | 文献类型 | 版本类型 | 开放类型 | 使用许可 | ||
fpls-10-00612.pdf(4260KB) | 期刊论文 | 出版稿 | 开放获取 | CC BY-NC-SA | 浏览 请求全文 |
除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。
修改评论