title: Development and application of West Nile virus subgenomic replicon RNA expressing secreted alkaline phosphatase.
abstract: We have developed a West Nile virus (WNV) subgenomic replicon harboring the secreted alkaline phosphatase (SEAP) reporter gene instead of viral structural genes (designated repWNV/SEAP). The repWNV/SEAP allowed easy evaluation of viral replication efficiency by direct measurement of SEAP secretion in the cell culture medium in physical containment level 2 facilities. Furthermore, we validated the availability of this system using a known anti-flavivirus gene, mouse oligoadenylate synthetase 1b (Oas1b). The Oas1b-transfected cells were more resistant to repWNV/SEAP replication than the original cells. Thus, this system not only affords a useful tool for identification/evaluation of anti-flavivirus genes/drugs in terms of safety, ease of use and reliability, but should be able to reduce or replace the bioassay using laboratory animals.
authors.affiliation: Laboratory of Laboratory Animal Science and Medicine, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita, Nishi, Kita-ku Sapporo, Hokkaido, Japan.
authors.forename: Kanako
authors.initials: K
authors.lastname: Moritoh
authors.forename: Akihiko
authors.initials: A
authors.lastname: Maeda
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Nishino
authors.forename: Nobuya
authors.initials: N
authors.lastname: Sasaki
authors.forename: Takashi
authors.initials: T
authors.lastname: Agui
mesh_terms: D015088:2',5'-Oligoadenylate Synthetase / Q000235:genetics / Q000378:metabolism; D000469:Alkaline Phosphatase; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D017930:Genes, Reporter; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism*; D012093:Replicon; D014764:Viral Proteins; D014902:West Nile virus / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
chemical_list: D012367:RNA, Viral; D014764:Viral Proteins; C509914:Oas1b protein, mouse; D015088:2',5'-Oligoadenylate Synthetase; D000469:Alkaline Phosphatase