title: In vitro inhibition of CSFV replication by multiple siRNA expression.
abstract: Classical swine fever (CSF) is a highly contagious viral disease of pigs which causes major economic losses worldwide. No specific drug is currently available for the effective treatment of CSFV infection; however, RNA interference (RNAi) has been applied successfully to inhibit the replication of human and other animal viruses. In this study, three effective siRNAs targeting NS3 of CSFV were selected. siNS3-2 targeting NS3 gene was chosen for further experimentation, while siN1 and siN2 targeting N(pro) gene, and siNS5B targeting NS5B gene describe previously. Single, double and quadruple anti-CSFV siRNA expression plasmids, with loxp sites at each end of the selectable marker genes, were constructed and analyzed using the same promoters or four different promoters, targeting N(pro), NS3 and NS5B genes of CSFV. Results indicate that single or multiple siRNA expression plasmids can efficiently inhibit CSFV replication and that inhibition was markedly stronger when multiple siRNAs were expressed targeting different genes of CSFV. Since RNAi applied to anti-CSFV research, this study provides anti-CSFV methods by single and multiple siRNA expression which can target most viral isolates of different subtypes and prevent viral escape. It also provides a basis for development of CSFV-resistant transgenic pigs.
authors.affiliation: Institute of Veterinary Sciences, Academy of Military Medical Sciences, 666 Liuying West Road, Changchun 130122, China.
authors.forename: Jiangnan
authors.initials: J
authors.lastname: Li
authors.forename: Yajuan
authors.initials: Y
authors.lastname: Dai
authors.forename: Shuai
authors.initials: S
authors.lastname: Liu
authors.forename: Huancheng
authors.initials: H
authors.lastname: Guo
authors.forename: Tiedong
authors.initials: T
authors.lastname: Wang
authors.forename: Hongsheng
authors.initials: H
authors.lastname: Ouyang
authors.forename: Changchun
authors.initials: C
authors.lastname: Tu
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006692:Classical Swine Fever Virus / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D017930:Genes, Reporter; D005814:Genes, Viral; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D013552:Swine; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D034741:RNA, Small Interfering; D017361:Viral Nonstructural Proteins