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Updated: Aug 30, 2026
High-throughput Antiviral Assays to Screen for Inhibitors of Zika Virus Replication
Published on: October 30, 2021
Potential high-throughput assay for screening inhibitors of West Nile virus replication
Michael K Lo1, Mark Tilgner, Pei-Yong Shi
1Wadsworth Center, New York State Department of Health, University at Albany, State University of New York, Albany, New York 12201, USA.
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.
Insights
A new West Nile virus (WNV) reporting cell line enables high-throughput screening for antiviral drugs. This tool accelerates the discovery of inhibitors targeting WNV and similar flaviviruses, crucial for public health.
Area of Science:
- Virology
- Molecular Biology
- Drug Discovery
Background:
- West Nile virus (WNV) and other flaviviruses pose significant public health threats.
- Effective prevention and treatment strategies are urgently needed.
- Current methods for screening antiviral compounds can be time-consuming.
Purpose of the Study:
- To develop a novel reporting cell line for high-throughput screening of WNV inhibitors.
- To establish a reliable assay for evaluating potential anti-WNV therapeutics.
- To demonstrate the applicability of this approach for other flaviviruses.
Main Methods:
- Engineered a WNV subgenomic replicon with dual reporter genes (Renilla luciferase and neomycin phosphotransferase) to create Rluc/NeoRep.
- Generated a stable BHK-21 cell line containing persistently replicating Rluc/NeoRep.
- Assessed inhibitor efficacy by measuring changes in Rluc activity and replicon RNA levels.
Main Results:
- The WNV reporting cell line demonstrated decreased Rluc activity and replicon RNA levels upon incubation with known WNV inhibitors.
- The assay's efficacy in measuring inhibitor potency was comparable to traditional viral infection assays.
- The developed cell line proved effective for high-throughput screening.
Conclusions:
- The WNV reporting cell line serves as a valuable high-throughput assay for anti-WNV drug discovery.
- This genetics-based antiviral assay approach is adaptable for other flaviviruses.
- The study provides a foundation for developing new treatments against flaviviral infections.
