Potential High-Throughput Assay for Screening Inhibitors of West Nile Virus Replication

Michael K. Lo, Mark Tilgner, Pei-Yong Shi

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Prevention and treatment of infection by West Nile virus (WNV) and other flaviviruses are public health priorities. We describe a reporting cell line that can be used for high-throughput screening of inhibitors against all targets involved in WNV replication. Dual reporter genes, encoding Renilla luciferase (Rluc) and neomycin phosphotransferase (Neo), were engineered into a WNV subgenomic replicon, resulting in Rluc/NeoRep. Geneticin selection of BHK-21 cells transfected with Rluc/NeoRep yielded a stable cell line that contains persistently replicating replicons. Incubation of the reporting cells with known WNV inhibitors decreased Rluc activity, as well as the replicon RNA level. The efficacies of the inhibitors, as measured by the depression of Rluc activity in the reporting cells, are comparable to those derived from authentic viral infection assays. Therefore, the WNV reporting cell line can be used as a high-throughput assay for anti-WNV drug discovery. A similar approach should be applicable to development of genetics-based antiviral assays for other flaviviruses.

Original languageEnglish (US)
Pages (from-to)12901-12906
Number of pages6
JournalJournal of Virology
Volume77
Issue number23
DOIs
StatePublished - Dec 2003
Externally publishedYes

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High-Throughput Screening Assays
Renilla Luciferases
West Nile virus
Virus Replication
virus replication
luciferase
screening
replicon
Replicon
assays
Flaviviridae
Flavivirus
cell lines
Cell Line
geneticin
Kanamycin Kinase
Health Priorities
kanamycin kinase
Virus Diseases
cells

ASJC Scopus subject areas

  • Immunology

Cite this

Potential High-Throughput Assay for Screening Inhibitors of West Nile Virus Replication. / Lo, Michael K.; Tilgner, Mark; Shi, Pei-Yong.

In: Journal of Virology, Vol. 77, No. 23, 12.2003, p. 12901-12906.

Research output: Contribution to journalArticle

Lo, Michael K. ; Tilgner, Mark ; Shi, Pei-Yong. / Potential High-Throughput Assay for Screening Inhibitors of West Nile Virus Replication. In: Journal of Virology. 2003 ; Vol. 77, No. 23. pp. 12901-12906.
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