{"doi":"10.1128/aac.49.12.4980-4988.2005","title":"High-Throughput Assays Using a Luciferase-Expressing Replicon, Virus-Like Particles, and Full-Length Virus for West Nile Virus Drug Discovery","abstract":"<jats:title>ABSTRACT</jats:title>\n          <jats:p>\n            Many flaviviruses cause significant human disease worldwide. The development of flavivirus chemotherapy requires reliable high-throughput screening (HTS) assays. Although genetic systems have been developed for many flaviviruses, their usage in antiviral HTS assays has not been well explored. Here we compare three cell-based HTS assays for West Nile virus (WNV) drug discovery: (i) an assay that uses a cell line harboring a persistently replicating subgenomic replicon (containing a deletion of viral structural genes), (ii) an assay that uses packaged virus-like particles containing replicon RNA, and (iii) an assay that uses a full-length reporting virus. A\n            <jats:italic>Renilla</jats:italic>\n            luciferase gene was engineered into the replicon or into the full-length viral genome to monitor viral replication. Potential inhibitors could be identified through suppression of luciferase signals upon compound incubation. The antiviral assays were optimized in a 96-well format, validated with known WNV inhibitors, and proved useful in identifying a new inhibitor(s) through HTS of a compound library. In addition, because each assay encompasses multiple but discrete steps of the viral life cycle, the three systems could potentially be used to discriminate the mode of action of any inhibitor among viral entry (detected by assays ii and iii but not by assay i), replication (including viral translation and RNA synthesis; detected by assays i to iii), and virion assembly (detected by assay iii but not by assays i and ii). The approaches described in this study should be applicable to the development of cell-based assays for other flaviviruses.\n          </jats:p>","journal":"Antimicrobial Agents and Chemotherapy","year":2005,"id":32144,"datarank":4.986834577216701,"base_score":4.762173934797756,"endowment":4.762173934797756,"self_citation_contribution":0.7143260902196635,"citation_network_contribution":4.272508486997038,"self_endowment_contribution":0.7143260902196635,"citer_contribution":4.272508486997038,"corpus_percentile":null,"corpus_rank":null,"citation_count":116,"citer_count":78,"citers_with_citation_signal":69,"citers_with_endowment":69,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":170738,"name":"Tia S. Deas","orcid":null,"position":1,"is_corresponding":false},{"id":170739,"name":"Ping Ren","orcid":null,"position":2,"is_corresponding":false},{"id":170740,"name":"Mark Tilgner","orcid":null,"position":3,"is_corresponding":false},{"id":12676,"name":"David M. Ferguson","orcid":"0000-0001-6294-8057","position":4,"is_corresponding":false},{"id":38203,"name":"Pei-Yong Shi","orcid":null,"position":5,"is_corresponding":false},{"id":170737,"name":"Francesc Puig-Basagoiti","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":4.762173934797756,"endowment":4.762173934797756,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"16304161","pmcid":"PMC1315944","openalex_id":"https://openalex.org/W2097289915","authors":[],"funders":[{"funder_name":"NIAID NIH HHS","grant_id":"U01 AI061193","title":null},{"funder_name":"NIAID NIH HHS","grant_id":"N01 AI025490","title":null},{"funder_name":"NIAID NIH HHS","grant_id":"AI061193","title":null},{"funder_name":"NIAID NIH HHS","grant_id":"AI065562","title":null},{"funder_name":"NIAID NIH HHS","grant_id":"R21 AI065562","title":null}],"total_grants":5,"fwci":7.5266,"citation_percentile":0.97151871,"influential_citations":5,"citation_trend":[{"year":2012,"count":2},{"year":2013,"count":5},{"year":2014,"count":11},{"year":2015,"count":1},{"year":2016,"count":4},{"year":2017,"count":7},{"year":2018,"count":3},{"year":2019,"count":2},{"year":2020,"count":12},{"year":2021,"count":2},{"year":2022,"count":6},{"year":2023,"count":1},{"year":2024,"count":2},{"year":2025,"count":3},{"year":2026,"count":1}],"oa_status":"green","license":"https://journals.asm.org/non-commercial-tdm-license","oa_locations":[{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/1315944","host_type":"repository"},{"url":"https://europepmc.org/articles/pmc1315944?pdf=render","host_type":"GREEN"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/1315944","host_type":"repository"},{"url":"https://journals.asm.org/doi/pdf/10.1128/AAC.49.12.4980-4988.2005","host_type":"publisher"},{"url":"https://doi.org/10.1128/aac.49.12.4980-4988.2005","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/16304161","host_type":"repository"}],"fields_of_study":["Mosquito-borne diseases and control","HIV Research and Treatment","Viral Infections and Vectors","Biology","Medicine","Antiviral Agents","Biological Assay","Drug Evaluation, Preclinical","Luciferases","Replicon","Virus Replication","West Nile virus"],"mesh_terms":["Antiviral Agents","Biological Assay","Drug Evaluation, Preclinical","Luciferases","Replicon","Virus Replication","West Nile virus"],"keywords":["Replicon","Flavivirus","Biology","Virology","Subgenomic mRNA","Viral replication","Virus","Luciferase","Antiviral drug","Drug discovery","High-throughput screening","Cell culture","RNA","Molecular biology","Gene","Transfection","Genetics","Bioinformatics","Genome"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-09T10:40:06.839408Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}