Suppression of bovine viral diarrhea virus replication by small interfering RNA and short hairpin RNA-mediated RNA interference.
Bovine viral diarrhea virus (BVDV) is a ubiquitous viral pathogen that affects cattle herds' worldwide causing significant economic loss. The current strategies to control BVDV infection include vaccination (modified-live or killed) and control of virus spread by enhanced biosecurity management, however, the disease remains prevalent. With the discovery of the sequence-specific method of gene silencing known as RNA interference (RNAi), a new era in antiviral therapies has begun. Here we report the efficient inhibition of BVDV replication by small interfering (siRNA) and short hairpin RNA (shRNA)-mediated gene silencing. siRNAs were generated to target the 5' non-translated (NTR) region and the regions encoding the C, NS4B and NS5A proteins of the BVDV genome. The siRNAs were first validated using an EGFP/BVDV reporter system and were then shown to suppress BVDV-induced cytopathic effects and viral titers in cell culture with surprisingly different activities compared to the reporter system. Efficient viral suppression was then achieved by bovine 7SK-expressed BVDV-specific shRNAs. Overall, our results demonstrated the use of siRNA and shRNA-mediated gene silencing to achieve efficient inhibition of the replication of this virus in cell culture.
Lambeth
Luke S
LS
CSIRO Livestock Industries, Australian Animal Health Laboratory, Private Bag 24, Geelong, Victoria 3220, Australia. ls.lambeth@gmail.com
Moore
Robert J
RJ
Muralitharan
Morley S
MS
Doran
Timothy J
TJ
D000818:Animals; D000998:Antiviral Agents; D001483:Base Sequence; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D002417:Cattle; D002460:Cell Line; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000502:physiology*; D020868:Gene Silencing; D015316:Genetic Therapy / Q000379:methods / Q000662:veterinary; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000502:physiology*; D012367:RNA, Viral / Q000096:biosynthesis*; D014162:Transfection; D014779:Virus Replication / Q000235:genetics* / Q000276:immunology
D000998:Antiviral Agents; D034741:RNA, Small Interfering; D012367:RNA, Viral