title: Bovine fetal epithelium cells expressing shRNA targeting viral VP1 gene resisted against foot-and-mouth disease virus.
abstract: 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.
authors.affiliation: Dairy Cattle Research Center, Shandong Academy of Agricultural Sciences, Jinan 250100, PR China.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Wang
authors.forename: Xiao
authors.initials: X
authors.lastname: Liu
authors.forename: Jianming
authors.initials: J
authors.lastname: Wu
authors.forename: Gang
authors.initials: G
authors.lastname: Wu
authors.forename: Li
authors.initials: L
authors.lastname: Yu
authors.forename: Chenqiang
authors.initials: C
authors.lastname: He
authors.forename: Huiting
authors.initials: H
authors.lastname: Yang
authors.forename: Weixin
authors.initials: W
authors.lastname: Xie
authors.forename: Xianzhu
authors.initials: X
authors.lastname: Xia
authors.forename: Hongbin
authors.initials: H
authors.lastname: He
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000998:Antiviral Agents / Q000378:metabolism*; D015152:Blotting, Northern; D015139:Blotting, Southern; D036022:Capsid Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D002417:Cattle; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000821:virology; D005347:Fibroblasts / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000254:growth & development*; D015870:Gene Expression; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D000998:Antiviral Agents; D036022:Capsid Proteins; D034741:RNA, Small Interfering; C442501:VP1 protein, Foot-and-mouth disease virus