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Highly efficient activation of endogenous gene in grape using CRISPR/dCas9-based transcriptional activators | |
Ren, Chong; Li, Huayang1; Liu, Yanfei1; Li, Shaohua; Liang, Zhenchang | |
2022 | |
发表期刊 | HORTICULTURE RESEARCH |
ISSN | 2662-6810 |
卷号 | 9 |
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. |
学科领域 | Plant Sciences ; Genetics & Heredity ; Horticulture |
DOI | 10.1093/hr/uhab037 |
收录类别 | SCI |
语种 | 英语 |
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) |
WOS记录号 | WOS:000791049400017 |
出版者 | OXFORD UNIV PRESS INC |
文献子类 | Article |
出版地 | CARY |
EISSN | 2052-7276 |
资助机构 | National Natural Science Foundation of China [31772266, 32001994] ; Bureau of International Cooperation of the Chinese Academy of Sciences [151111KYSB20170032] |
作者邮箱 | zl249@ibca.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/2S10CLM1/28792 |
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, Beijing Key Lab Grape Sci & Enol, Key Lab Plant Resource, 20 Nanxincun, Beijing 100093, Peoples R China 2.Univ Chinese Acad Sci, 19 Yuquan Rd, Beijing 100049, 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). |
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