In vitro RNA interference targeting the DNA polymerase gene inhibits orf virus replication in primary ovine fetal turbinate cells

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...

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Bibliographic Details
Published in:Archives of virology 2014-05, Vol.159 (5), p.915-920
Main Authors: Wang, Gaili, He, Wenqi, Song, Deguang, Li, Jida, Bao, Yingfu, Lu, Rongguang, Bi, Jingying, Zhao, Kui, Gao, Feng
Format: Article
Language:English
Subjects:
Animals
antiviral properties
Bioengineering
Biomedical and Life Sciences
Biomedicine
Cell culture
Cells, Cultured
cytology
cytopathogenicity
DNA polymerase
DNA-directed DNA polymerase
DNA-Directed DNA Polymerase - genetics
DNA-Directed DNA Polymerase - metabolism
Genes
Genetic testing
genetics
Genomes
Genomics
Infections
Infectious Diseases
Medical Microbiology
metabolism
Orf virus
Orf virus - genetics
Orf virus - physiology
Original
Original Article
physiology
quantitative polymerase chain reaction
RNA interference
RNA Interference - physiology
Sheep
small interfering RNA
titration
Turbinates
Turbinates - cytology
Virology
Virus Cultivation
virus replication
Virus Replication - genetics
Virus Replication - physiology
viruses
Zoonoses
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Summary: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.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-013-1896-z