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Cooperative antivirus activities of two duplicated viperin homeologs confirmed by CRISPR/Cas9 editing in hexaploid gibel carp

文献类型： 外文期刊

作者： Mou, Cheng-Yan 1 ; Wang, Yang 1 ; Yu, Peng 1 ; Li, Zhi 1 ; Wang, Zhong-Wei 1 ; Li, Xi-Yin 1 ; Li, Shun 1 ; Lu, Long-Feng 1 ; Tong, Jin-Feng 1 ; Zhang, Qi-Ya 1 ; Zhang, Xiao-Juan 1 ; Zhou, Li 1 ; Gui, Jian-Fang 1 ;

作者机构： 1.Chinese Acad Sci, Innovat Acad Seed Design, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China

2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China

3.Hubei Hongshan Lab, Wuhan 430070, Peoples R China

4.Sichuan Acad Agr Sci, Fisheries Inst, Chengdu 611731, Sichuan, Peoples R China

关键词： Viperin; Homeolog; Gibel carp; Herpesvirus; CRISPR/Cas9

期刊名称：AQUACULTURE （ 影响因子：5.135； 五年影响因子：5.125 ）

ISSN： 0044-8486

年卷期： 2022 年 548 卷

页码：

收录情况： SCI

摘要： Viperin is increasingly thought as a key protein in antiviral response. However, the in vivo antiviral effects of fish viperin remains unresolved. Hexaploid gibel carp (Carassius gibelio) is an important aquaculture species in China and its culture industry has suffered enormous economic losses due to an epizootic disease caused by C. auratus herpesvirus (CaHV) since 2012. In our previous studies, two duplicated viperin homeologs (Cgviperin-A and Cgviperin-B) were identified and their divergent antiviral mechanisms were revealed through in vitro overexpression in CAB (C. auratus blastulae embryonic) cell. Here, we tried to explore their function difference in vivo by using CRISPR/Cas9 editing. Firstly, we uncovered their biased expression patterns and differential promoter activities. Then we mutated them singly or simultaneously in resistant clone H of gibel carp by using CRISPR/ Cas9 and finally confirmed that they cooperatively participated in the antiviral battle between gibel carp and CaHV through virus challenge and comparative transcriptome analysis. CgViperin-A depletion mainly affected immune-related pathways and Cgviperin-B deficiency influenced autophagy-associated pathways. Simultaneous disruption of them resulted in complete loss of resistance ability of gibel carp against CaHV. Significantly, the deficiency of Cgviperin-A influenced the expression of Cgviperin-B in Cgviperin-A(-/-/-) mutant, but Cgviperin-A transcripts kept a high level of expression in the Cgviperin-B-/-/- mutant, same as in the WT. Therefore, the current findings are not only the first time to clarify the antiviral activities of Viperin in the battle between teleost and virus in vivo, but also explain the detailed evolutionary divergence of a duplicated gene in a recurrent polyploid fish.

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