Short hairpin RNA-mediated silencing of bovine rotavirus NSP4 gene prevents diarrhoea in suckling mice
文献类型： 外文期刊
作者： Chen, Fangyuan 1 ; Wang, Hongmei 1 ; He, Hongbin 1 ; Song, Lingling 1 ; Wu, Jianming 1 ; Gao, Yundong 1 ; Liu, Xiao 1 ; H 1 ;
作者机构： 1.Shandong Acad Agr Sci, Dairy Cattle Res Ctr, Jinan 250100, Peoples R China
2.NE Agr Univ, Coll Life Sci, Harbir 150030, Peoples R China
3.Shandong Normal Univ, Coll Life Sci, Jinan 250014, Peoples R China
4.Inner Mongolia Univ, Coll Life Sci, Hohhot 010021, Peoples R China
5.Univ Penn, Div G
期刊名称：JOURNAL OF GENERAL VIROLOGY （ 影响因子：3.891； 五年影响因子：3.719 ）
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收录情况： SCI
摘要： While RNA interference (RNAi) has been widely used to study rotavirus gene function in vitro, the potential therapeutic role for RNAi in vivo has not been explored. To this end, we constructed two recombinant lentiviral vectors containing short hairpin RNA (shRNA) against non-structural protein-4 (NSP4) of bovine rotavirus (BRV), RNAi-351 and RNAi-492. RNAi-351 and RNAi-492 strongly suppressed the transient expression of a FLAG-tagged NSP4 fusion protein in 293T cells. In BRV-susceptible MA104 cells, RNAi-492 more potently silenced NSP4 mRNA than RNAi-351 and combination of the two shRNAs almost completely silenced viral NSP4 gene expression. While 100% of suckling mice exposed to BRV and control shRNA developed severe diarrhoea, no suckling mice exposed to BRV in the presence of RNAi-492 or a combination of RNAi-492/RNAi-351 developed severe diarrhoea, and only 20 and 3.3% developed mild diarrhoea, respectively. In addition, RNAi-492 and RNAi-351 markedly abrogated rotaviral replication in MA104 cells and significantly inhibited BRV replication in mouse pups. These results indicated that shRNAs silencing NSP4 gene had substantial antiviral properties and inhibited replication of BRV in a sequence-specific manner that may have clinical application.
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