Suppression of porcine reproductive and respiratory syndrome virus replication in MARC-145 cells by shRNA targeting ORF1 region.
Porcine reproductive and respiratory syndrome (PRRS) is an economically important disease in swine producing area. The current vaccine strategies cannot provide complete protection against PRRSV. The objective of this study was to determine if specific short-hairpin RNA (shRNA) directed against different genomic regions of ORF1b of PRRSV could be utilized to inhibit virus replication in MARC-145 cells. Two shRNA expression vectors targeting ORF1b gene of PRRSV were constructed and delivered into MARC-145 cells, and then infected with PRRSV. The results showed that PRRSV-specific cytopathic effect (CPE) could be inhibited in the cells transfected with pSUPER-P2 and pSUPER-P3, and the virus titers in the cells transfected with pSUPER-P2 and pSUPER-P3 were lower than those control cells by approximately 100 fold. Moreover, the expression of ORF1 of PRRSV in the cells was reduced both at RNA and protein levels comparing to the controls. It indicated that vector-based shRNA targeting ORF1 region could effectively inhibit PRRSV replication in MARC-145 cells.
Key Laboratory of Animal Diseases Diagnostic and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agriculture University, Nanjing 210095, PR China.
Guanming
G
Li
Juan
J
Huang
Ping
P
Jiang
Yufeng
Y
Li
Wenming
W
Jiang
Xianwei
X
Wang
D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism*; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral; D020868:Gene Silencing; D005814:Genes, Viral; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics* / Q000254:growth & development*; D034741:RNA, Small Interfering / Q000378:metabolism*; D013552:Swine; D014779:Virus Replication
D000998:Antiviral Agents; D034741:RNA, Small Interfering