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Journal of Virology, December 2003, p. 12901-12906, Vol. 77, No. 23
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.23.12901-12906.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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

Michael K. Lo,1 Mark Tilgner,1 and Pei-Yong Shi1,2*

Wadsworth Center, New York State Department of Health,1 Department of Biomedical Sciences, University at Albany, State University of New York, Albany, New York 122012

Received 9 June 2003/ Accepted 22 August 2003

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.

* Corresponding author. Mailing address: Wadsworth Center, New York State Department of Health, Albany, NY 12201. Phone: (518) 473-7487. Fax: (518) 473-1326. E-mail: ship{at}wadsworth.org.

Journal of Virology, December 2003, p. 12901-12906, Vol. 77, No. 23
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.23.12901-12906.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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