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Highly efficient activation of endogenous gene in grape using CRISPR/dCas9-based transcriptional activators

文献类型：期刊论文

作者Ren, Chong; Li, Huayang2; Liu, Yanfei2; Li, Shaohua; Liang, Zhenchang
刊名HORTICULTURE RESEARCH
出版日期2022
卷号9
ISSN号2662-6810
DOI10.1093/hr/uhab037
文献子类Article
英文摘要Overexpression and knockout (or knockdown) of gene of interest are two commonly used strategies for gene functional study. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system-mediated gene knockout has been applied in most plant species, including grapevine. However, CRISPR/dCas9 (deactivated Cas9)-based transcriptional activation is still unreported in fruit crops, although a few studies have been documented in Arabidopsis and rice. Here, we tested two transcriptional activators VP64 and TV for transcriptional activation of endogenous genes in grape. Both the dCas9-VP64 and dCas9-TV systems are efficient enough for transcriptional activation of the UDP-glucose flavonoid glycosyltransferases (UFGT) gene in grape cells. The effectiveness of the dCas9-VP64 system in UFGT activation was about 1.6- to 5.6-fold, while the efficiency of the dCas9-TV system was around 5.7- to 7.2-fold. Moreover, in grapevine plants, highly efficient activation of the cold-responsive transcription factor gene CBF4 was achieved by using the dCas9-TV system. The expression of CBF4 was increased 3.7- to 42.3-fold in transgenic plants. Compared with the wild-type plants, the CBF4-activated plants exhibited lower electrolyte leakage after cold treatment. Our results demonstrate the effectiveness of the dCas9-VP64 and dCas9-TV systems in gene activation in grape, which will facilitate application of transcriptional activation in this economically important species.
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VITIS-VINIFERA ; TARGETED MUTAGENESIS ; CRISPR/CAS9 SYSTEM ; COLD-ACCLIMATION ; LOW-TEMPERATURE ; FAMILY ; PLANT ; EXPRESSION ; TOOLBOX
WOS研究方向Science Citation Index Expanded (SCI-EXPANDED)
语种英语
WOS记录号WOS:000791049400017
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/2S10CLM1/28792]  
专题中科院北方资源植物重点实验室
作者单位1.Univ Chinese Acad Sci, 19 Yuquan Rd, Beijing 100049, Peoples R China
2.Chinese Acad Sci, Inst Bot, Beijing Key Lab Grape Sci & Enol, Key Lab Plant Resource, 20 Nanxincun, Beijing 100093, Peoples R China
推荐引用方式
GB/T 7714		 Ren, Chong,Li, Huayang,Liu, Yanfei,et al. Highly efficient activation of endogenous gene in grape using CRISPR/dCas9-based transcriptional activators[J]. HORTICULTURE RESEARCH,2022,9.
APA Ren, Chong,Li, Huayang,Liu, Yanfei,Li, Shaohua,&Liang, Zhenchang.(2022).Highly efficient activation of endogenous gene in grape using CRISPR/dCas9-based transcriptional activators.HORTICULTURE RESEARCH,9.
MLA Ren, Chong,et al."Highly efficient activation of endogenous gene in grape using CRISPR/dCas9-based transcriptional activators".HORTICULTURE RESEARCH 9(2022).

入库方式： OAI收割

来源：植物研究所

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