title: In vitro RNA interference targeting the DNA polymerase gene inhibits orf virus replication in primary ovine fetal turbinate cells.
abstract: Orf, which is caused by orf virus (ORFV), is distributed worldwide and is endemic in most sheep- and/or goat-raising countries. RNA interference (RNAi) pathways have emerged as important regulators of virus-host cell interactions. In this study, the specific effect of RNAi on the replication of ORFV was explored. The application of RNA interference (RNAi) inhibited the replication of ORFV in cell culture by targeting the ORF025 gene of ORFV, which encodes the viral polymerase. Three small interfering RNA (siRNA) (named siRNA704, siRNA1017 and siRNA1388) were prepared by in vitro transcription. The siRNAs were evaluated for antiviral activity against the ORFV Jilin isolate by the observation of cytopathic effects (CPE), virus titration, and real-time PCR. After 48 h of infection, siRNA704, siRNA1017 and siRNA1388 reduced virus titers by 59- to 199-fold and reduced the level of viral replication by 73-89 %. These results suggest that these three siRNAs can efficiently inhibit ORFV genome replication and infectious virus production. RNAi targeting of the DNA polymerase gene is therefore potentially useful for studying the replication of ORFV and may have potential therapeutic applications.
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun, 130062, China.
authors.forename: Gaili
authors.initials: G
authors.lastname: Wang
authors.forename: Wenqi
authors.initials: W
authors.lastname: He
authors.forename: Deguang
authors.initials: D
authors.lastname: Song
authors.forename: Jida
authors.initials: J
authors.lastname: Li
authors.forename: Yingfu
authors.initials: Y
authors.lastname: Bao
authors.forename: Rongguang
authors.initials: R
authors.lastname: Lu
authors.forename: Jingying
authors.initials: J
authors.lastname: Bi
authors.forename: Kui
authors.initials: K
authors.lastname: Zhao
authors.forename: Feng
authors.initials: F
authors.lastname: Gao
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D009923:Orf virus / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference / Q000502:physiology*; D012756:Sheep; D014420:Turbinates / Q000166:cytology*; D014776:Virus Cultivation; D014779:Virus Replication / Q000235:genetics / Q000502:physiology*
chemical_list: D004259:DNA-Directed DNA Polymerase