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
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spelling Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral ScreeningÁlvarez de Miranda Rodríguez, Francisco JavierMartín, RocíoAlcamí, AntonioHernáez, BrunoPoxvirusFluorescent reporterMpoxAntiviralClade IIbBisbenzimideThe 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.This research was funded by the Spanish Ministry of Science and Innovation grants PID2022-136867NB-I00 and RTI2021-128580OB-100 to B.H. and A.A., respectively, and by a La Caixa Foundation grant HR23-00558 to A.A.Peer reviewedMultidisciplinary Digital Publishing InstituteMinisterio de Ciencia e Innovación (España)Agencia Estatal de Investigación (España)Fundación la CaixaConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2025202520252025info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/382502reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2022-136867NB-I00info:eu-repo/grantAgreement/AEI//RTI2021-128580OB-100The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.3390/v17020253https://doi.org/10.3390/v17020253Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3825022026-05-22T06:33:51Z
dc.title.none.fl_str_mv Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening
title Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening
spellingShingle Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening
Álvarez de Miranda Rodríguez, Francisco Javier
Poxvirus
Fluorescent reporter
Mpox
Antiviral
Clade IIb
Bisbenzimide
title_short Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening
title_full Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening
title_fullStr Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening
title_full_unstemmed Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening
title_sort Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening
dc.creator.none.fl_str_mv Álvarez de Miranda Rodríguez, Francisco Javier
Martín, Rocío
Alcamí, Antonio
Hernáez, Bruno
author Álvarez de Miranda Rodríguez, Francisco Javier
author_facet Álvarez de Miranda Rodríguez, Francisco Javier
Martín, Rocío
Alcamí, Antonio
Hernáez, Bruno
author_role author
author2 Martín, Rocío
Alcamí, Antonio
Hernáez, Bruno
author2_role author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia e Innovación (España)
Agencia Estatal de Investigación (España)
Fundación la Caixa
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Poxvirus
Fluorescent reporter
Mpox
Antiviral
Clade IIb
Bisbenzimide
topic Poxvirus
Fluorescent reporter
Mpox
Antiviral
Clade IIb
Bisbenzimide
description 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.
publishDate 2025
dc.date.none.fl_str_mv 2025
2025
2025
2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/382502
url http://hdl.handle.net/10261/382502
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2022-136867NB-I00
info:eu-repo/grantAgreement/AEI//RTI2021-128580OB-100
The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.3390/v17020253
https://doi.org/10.3390/v17020253
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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