Bovine fetal epithelium cells expressing shRNA targeting viral VP1 gene resisted against foot-and-mouth disease virus.
Bovine fetal epithelium cells expressing shRNA targeting viral VP1 gene resisted against foot-and-mouth disease virus.
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DOI：
10.1016/j.virol.2013.02.003
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发表时间：
2013-05
期刊：
影响因子：
3.7
通讯作者：
Hongmei Wang;Xiao Liu;Jianming Wu;Gang Wu;Li Yu;Chenqiang He;Hui-Ling Yang;Weixin Xie;X. Xia;Hongbin He
中科院分区：
文献类型：
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作者：
Hongmei Wang;Xiao Liu;Jianming Wu;Gang Wu;Li Yu;Chenqiang He;Hui-Ling Yang;Weixin Xie;X. Xia;Hongbin He
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 TCID50of 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.
