title: Effective inhibition of porcine transmissible gastroenteritis virus replication in ST cells by shRNAs targeting RNA-dependent RNA polymerase gene.
abstract: Transmissible gastroenteritis virus (TGEV) is identified as one of the most important pathogenic agents during swine enteric infection, leading to high mortality in neonatal pigs and severe annual economic loss in swine-producing areas. Up to date, various vaccines developed against TGEV still need to be improved. To exploit the possibility of using RNA interference (RNAi) as a strategy against TGEV infection, two shRNA-expressing plasmids (pEGFP-U6/P1 and pEGFP-U6/P2) targeting the RNA-dependent RNA polymerase (RdRp) gene of TGEV were constructed and transfected into swine testicular (ST) cells. The cytopathic effect (CPE) and MTS assays demonstrated that both shRNAs were capable of protecting cells against TGEV invasion with very high specificity and efficiency. A real-time quantitative RT-PCR further confirmed that the amounts of viral RNAs in cell cultures pre-transfected with the two plasmids were reduced by 95.2% and up to 100%, respectively. Our results suggest that RNAi might be a promising new strategy against TGEV infection.
authors.affiliation: Department of Animal Science, School of Agriculture and Biology, Shanghai Jiao Tong University, 2678 Qixin Road, Shanghai 201101, China.
authors.forename: Jun-fang
authors.initials: JF
authors.lastname: Zhou
authors.forename: Xiu-guo
authors.initials: XG
authors.lastname: Hua
authors.forename: Li
authors.initials: L
authors.lastname: Cui
authors.forename: Jian-guo
authors.initials: JG
authors.lastname: Zhu
authors.forename: De-nian
authors.initials: DN
authors.lastname: Miao
authors.forename: Yong
authors.initials: Y
authors.lastname: Zou
authors.forename: Xi-zhong
authors.initials: XZ
authors.lastname: He
authors.forename: Wan-guo
authors.initials: WG
authors.lastname: Su
mesh_terms: D000818:Animals; D002460:Cell Line / Q000821:virology; D005761:Gastroenteritis, Transmissible, of Swine / Q000628:therapy / Q000821:virology*; D005814:Genes, Viral; D010957:Plasmids; D034622:RNA Interference; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics*; D012680:Sensitivity and Specificity; D013552:Swine; D014162:Transfection / Q000379:methods; D005760:Transmissible gastroenteritis virus / Q000502:physiology*; D014779:Virus Replication / Q000235:genetics
chemical_list: D012324:RNA-Dependent RNA Polymerase