Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening

The ongoing global outbreak of mpox caused by clade IIb viruses has led to more than 100,000 confirmed cases around the world, highlighting the urgent need for antiviral research to combat current and future mpox outbreaks. Reporter viruses expressing fluorescent proteins to monitor viral replicatio...

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Detalles Bibliográficos
Autores: Álvarez de Miranda Rodríguez, Francisco Javier, Martín, Rocío, Alcamí, Antonio, Hernáez, Bruno
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2025
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/382502
Acceso en línea:http://hdl.handle.net/10261/382502
Access Level:acceso abierto
Palabra clave:Poxvirus
Fluorescent reporter
Mpox
Antiviral
Clade IIb
Bisbenzimide
Descripción
Sumario:The ongoing global outbreak of mpox caused by clade IIb viruses has led to more than 100,000 confirmed cases around the world, highlighting the urgent need for antiviral research to combat current and future mpox outbreaks. Reporter viruses expressing fluorescent proteins to monitor viral replication and virus spreading in cell culture provide a powerful tool for antiviral drug screening. In this work, we engineered two recombinant mpox clade IIb viruses by inserting, under the control of the vaccinia early/late promoter 7.5, the coding sequence of two different fluorescent proteins (EGFP and TurboFP635) in a previously unreported location within the viral genome. These recombinant viruses replicate in BSC-1 cells at rates similar to those of the parental virus. We show how these reporter mpox viruses allow the discrimination of infected cells by cell flow cytometry and facilitate the quantification of viral spread in cell culture. Finally, we validated these reporter viruses with two previously known inhibitors of poxvirus replication, cytosine arabinoside (AraC) and bisbenzimide.