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

doi:10.1093/hr/uhab037

CORC > 植物研究所 > 中国科学院植物研究所 > 中科院北方资源植物重点实验室

	Highly efficient activation of endogenous gene in grape using CRISPR/dCas9-based transcriptional activators
	Ren, Chong ; Li, Huayang 2; Liu, Yanfei 2; Li, Shaohua ; Liang, Zhenchang
刊名	HORTICULTURE RESEARCH
	2022
卷号	9
ISSN号	2662-6810
DOI	10.1093/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
电子版国际标准刊号	2052-7276
出版地	CARY
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
出版者	OXFORD UNIV PRESS INC
WOS记录号	WOS:000791049400017
资助机构	National Natural Science Foundation of China [31772266, 32001994] ; Bureau of International Cooperation of the Chinese Academy of Sciences [151111KYSB20170032]
内容类型	期刊论文
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).