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Optimizing the CRISPR/Cas9 system for genome editing in grape by using grape promoters

文献类型：期刊论文

作者Ren, Chong1; Liu, Yanfei1,2; Guo, Yuchen1,2; Duan, Wei1; Fan, Peige1; Li, Shaohua1; Liang, Zhenchang1
刊名HORTICULTURE RESEARCH
出版日期2021
卷号8期号:1
ISSN号2662-6810
DOI10.1038/s41438-021-00489-z
文献子类Article
英文摘要The efficacy of the CRISPR/Cas9 system in grapevine (Vitis vinifera L.) has been documented, but the optimization of this system, as well as CRISPR/Cas9-mediated multiplex genome editing, has not been explored in this species. Herein, we identified four VvU3 and VvU6 promoters and two ubiquitin (UBQ) promoters in grapevine and demonstrated that the use of the identified VvU3/U6 and UBQ2 promoters could significantly increase the editing efficiency in grape by improving the expression of sgRNA and Cas9, respectively. Furthermore, we conducted multiplex genome editing using the optimized CRISPR/Cas9 vector that contained the conventional multiple sgRNA expression cassettes or the polycistronic tRNA-sgRNA cassette (PTG) by targeting the sugar-related tonoplastic monosaccharide transporter (TMT) family members TMT1 and TMT2, and the overall editing efficiencies were higher than 10%. The simultaneous editing of TMT1 and TMT2 resulted in reduced sugar levels, which indicated the role of these two genes in sugar accumulation in grapes. Moreover, the activities of the VvU3, VvU6, and UBQ2 promoters in tobacco genome editing were demonstrated by editing the phytoene desaturase (PDS) gene in Nicotiana benthamiana leaves. Our study provides materials for the optimization of the CRISPR/Cas9 system. To our knowledge, our simultaneous editing of the grape TMT family genes TMT1 and TMT2 constitutes the first example of multiplex genome editing in grape. The multiplex editing systems described in this manuscript expand the toolbox of grape genome editing, which would facilitate basic research and molecular breeding in grapevine.
学科主题Plant Sciences ; Genetics & Heredity ; Horticulture
出版地CARY
电子版国际标准刊号2052-7276
WOS关键词TARGETED MUTAGENESIS ; SUGAR ACCUMULATION ; CRISPR-CAS9 ; WHEAT ; DESIGN ; RICE
WOS研究方向Science Citation Index Expanded (SCI-EXPANDED)
语种英语
WOS记录号WOS:000624975000015
出版者OXFORD UNIV PRESS INC
资助机构National Natural Science Foundation of China [31772266, 32001994] ; Bureau of International Cooperation of the Chinese Academy of Sciences [151111KYSB20170032]
源URL[http://ir.ibcas.ac.cn/handle/2S10CLM1/26650]  
专题中科院北方资源植物重点实验室
作者单位1.Chinese Acad Sci, Innovat Acad Seed Design, Inst Bot, Beijing Key Lab Grape Sci & Enol, Beijing 100093, Peoples R China
2.Chinese Acad Sci, Innovat Acad Seed Design, Inst Bot, CAS Key Lab Plant Resources, Beijing 100093, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式
GB/T 7714		 Ren, Chong,Liu, Yanfei,Guo, Yuchen,et al. Optimizing the CRISPR/Cas9 system for genome editing in grape by using grape promoters[J]. HORTICULTURE RESEARCH,2021,8(1).
APA Ren, Chong.,Liu, Yanfei.,Guo, Yuchen.,Duan, Wei.,Fan, Peige.,...&Liang, Zhenchang.(2021).Optimizing the CRISPR/Cas9 system for genome editing in grape by using grape promoters.HORTICULTURE RESEARCH,8(1).
MLA Ren, Chong,et al."Optimizing the CRISPR/Cas9 system for genome editing in grape by using grape promoters".HORTICULTURE RESEARCH 8.1(2021).

入库方式： OAI收割

来源：植物研究所

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