Development of a Synthetic Genomics-Based Venezuelan Equine Encephalitis Virus Expressing Secreted Gaussia Luciferase for High-Throughput Antiviral Screening
- 1Laboratory for Viral Infection Control, Frontier Science Research Center, University of Miyazaki
Abstract
Venezuelan equine encephalitis virus (VEEV) is a mosquito-borne alphavirus that causes severe neurological disease in humans and animals. Despite its public health importance, no approved antiviral therapy is currently available, and conventional methods for quantifying infectious virus are labor-intensive and poorly suited for high-throughput antiviral screening. Furthermore, research on VEEV is complicated by the restricted availability and international transport of infectious viral materials. Here, we established a full-length infectious cDNA clone of VEEV entirely from chemically synthesized DNA fragments and rescued recombinant reporter viruses expressing mCherry2, ZsGreen, or the secreted Gaussia luciferase mutant 23 (GLucM23). The reporter viruses retained replication kinetics comparable to those of wild-type VEEV, while the fluorescent reporters enabled direct visualization of infection and virus-induced cytopathic effects. GLucM23-expressing VEEV allowed rapid, sensitive, and non-destructive quantification of viral replication directly from culture supernatants, enabling repeated longitudinal measurements without disrupting infected cells. We further demonstrated the utility of this system by screening approximately 3,000 FDA-approved compounds to identify inhibitors of VEEV replication. Together, these findings establish a versatile synthetic genomics-based reporter virus platform for quantitative studies of VEEV replication and scalable antiviral drug discovery.
Related
- The increased production of monocytes with immunosuppressive transcriptional profiles predicts the development of SIV encephalitis
- Development of a high throughput continuous fluorescence anisotropy-based assay to analyse peptidoglycan-digesting enzymes
- Gene transcriptional profiling of osmoregulatory stress response in salmonids: Development and assessment of high-throughput multispecies assays
- Transgenic Plasmodium berghei expressing P. vivax pre-erythrocytic antigens SSP3 and SPECT1 to elucidate their efficacy towards vaccine development
- Multidimensional analysis of social discourse on Twitter (X) around Venezuelan migration in Colombia
BibTeX
@misc{kawano2026development,
title = {{Development of a Synthetic Genomics-Based Venezuelan Equine Encephalitis Virus Expressing Secreted Gaussia Luciferase for High-Throughput Antiviral Screening}},
author = {M. X. Kawano and A. Ohkawa and N. Suwongsaksri and A. Saito},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.21.760855},
url = {https://www.biorxiv.org/content/10.64898/2026.10.21.760855v1},
note = {CC BY-NC},
}
RIS
TY - UNPB
TI - Development of a Synthetic Genomics-Based Venezuelan Equine Encephalitis Virus Expressing Secreted Gaussia Luciferase for High-Throughput Antiviral Screening
AU - M. X. Kawano
AU - A. Ohkawa
AU - N. Suwongsaksri
AU - A. Saito
PY - 2026
DA - 2026/10/21
PB - bioRxiv
DO - 10.64898/2026.10.21.760855
UR - https://www.biorxiv.org/content/10.64898/2026.10.21.760855v1
AB - Venezuelan equine encephalitis virus (VEEV) is a mosquito-borne alphavirus that causes severe neurological disease in humans and animals. Despite its public health importance, no approved antiviral therapy is currently available, and conventional methods for quantifying infectious virus are labor-intensive and poorly suited for high-throughput antiviral screening. Furthermore, research on VEEV is complicated by the restricted availability and international transport of infectious viral materials. Here, we established a full-length infectious cDNA clone of VEEV entirely from chemically synthesized DNA fragments and rescued recombinant reporter viruses expressing mCherry2, ZsGreen, or the secreted Gaussia luciferase mutant 23 (GLucM23). The reporter viruses retained replication kinetics comparable to those of wild-type VEEV, while the fluorescent reporters enabled direct visualization of infection and virus-induced cytopathic effects. GLucM23-expressing VEEV allowed rapid, sensitive, and non-destructive quantification of viral replication directly from culture supernatants, enabling repeated longitudinal measurements without disrupting infected cells. We further demonstrated the utility of this system by screening approximately 3,000 FDA-approved compounds to identify inhibitors of VEEV replication. Together, these findings establish a versatile synthetic genomics-based reporter virus platform for quantitative studies of VEEV replication and scalable antiviral drug discovery.
ER -