Effective inhibition of porcine transmissible gastroenteritis virus replication in ST cells by shRNAs targeting RNA-dependent RNA polymerase gene.
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.
Zhou
Jun-fang
JF
Department of Animal Science, School of Agriculture and Biology, Shanghai Jiao Tong University, 2678 Qixin Road, Shanghai 201101, China.
Hua
Xiu-guo
XG
Cui
Li
L
Zhu
Jian-guo
JG
Miao
De-nian
DN
Zou
Yong
Y
He
Xi-zhong
XZ
Su
Wan-guo
WG
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
D012324:RNA-Dependent RNA Polymerase