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Bovine fetal epithelium cells expressing shRNA targeting viral VP1 gene resisted against foot-and-mouth disease virus

文献类型： 外文期刊

作者： Wang, Hongmei 1 ; Liu, Xiao 1 ; Wu, Jianming 1 ; Wu, Gang 1 ; Yu, Li 2 ; He, Chenqiang 3 ; Yang, Huiting 1 ; Xie, Weixin; 1 ;

作者机构： 1.Shandong Acad Agr Sci, Dairy Cattle Res Ctr, Jinan 250100, Peoples R China

2.Harbin Vet Res Inst, State Key Lab Vet Biotechnol, Div Livestock Infect Dis, Harbin 150001, Peoples R China

3.Shandong Normal Univ, Coll Life Sci, Jinan 250014, Peoples R China

4.Acad Mil Med Sci, Inst Mil Vet, Key Lab Jinlin Prov Zoonosis Prevent & Control, Changchun 130122, Peoples R China

关键词： FMDV;RNAi;Transgenic bovine fetus

期刊名称：VIROLOGY （ 影响因子：3.616； 五年影响因子：3.967 ）

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收录情况： SCI

摘要： RNAi could protect experimental animals from Foot-and-mouth disease virus (FMDV), but pivotal issue is delivery of RNAi. In this study, shRNA recombinant lentiviral plasmid RNAi-LT6 targeting VP1 of FMDV, which strongly suppressed the transient expression of a FLAG-tagged VP1 protein in 293T cells and significantly inhibited viral replication in BHK-21 cells, was screened and transfected into bovine fetal fibroblast cells. With subsequent somatic cell cloning, three 4-month-old transgenic fetuses were obtained, and integration into chromosome and expression of shRNA in primary transgenic tongue epithelium cells were confirmed by Southern hybridization and Northern assay, respectively. shRNA significantly suppressed viral RNA synthesis and viral replication in primary transgenic tongue epithelium cells after inoculation of 200 TCID50 of FMDV. The results suggested that transgenic cloning may be a useful tool for RNAi delivery and RNAi anti-viral strategy, and RNAi-LT6 could be a candidate shRNA used for preparation of transgenic cattle against FMDV.

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