Potential High-Throughput Assay for Screening Inhibitors of West Nile Virus Replication
AUTOR(ES)
Lo, Michael K.
FONTE
American Society for Microbiology
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=262590Documentos Relacionados
- High-Throughput Detection of West Nile Virus RNA
- A three-fluorophore FRET assay for high-throughput screening of small-molecule inhibitors of ribosome assembly
- A high-throughput screening assay for distinguishing nitrile hydratases from nitrilases
- Fluorescence Detection-Based Functional Assay for High-Throughput Screening for MraY
- Model System for High-Throughput Screening of Novel Human Immunodeficiency Virus Protease Inhibitors in Escherichia coli
