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dc.creatorArias Arias, Jorge Luis
dc.creatorMacPherson, Derek J.
dc.creatorHill, Maureen E.
dc.creatorHardy, Jeanne A.
dc.creatorMora Rodríguez, Rodrigo Antonio
dc.date.accessioned2022-07-19T21:41:27Z
dc.date.available2022-07-19T21:41:27Z
dc.date.issued2020-01-09
dc.identifier.citationhttps://www.jbc.org/es_ES
dc.identifier.issn0021-9258
dc.identifier.urihttps://hdl.handle.net/10669/86985
dc.description.abstractThe genus Flavivirus in the family Flaviviridae comprises many medically important viruses, such as dengue virus (DENV), Zika virus (ZIKV), and yellow fever virus. The quest for thera- peutic targets to combat flavivirus infections requires a better understanding of the kinetics of virus–host interactions during infections with native viral strains. However, this is precluded by limitations of current cell-based systems for monitoring flavivi- rus infection in living cells. In the present study, we report the construction of fluorescence-activatable sensors to detect the activities of flavivirus NS2B–NS3 serine proteases in living cells. The system consists of GFP-based reporters that become fluo- rescent upon cleavage by recombinant DENV-2/ZIKV proteases in vitro. A version of this sensor containing the flavivirus inter- nal NS3 cleavage site linker reported the highest fluorescence activation in stably transduced mammalian cells upon DENV-2/ ZIKV infection. Moreover, the onset of fluorescence correlated with viral protease activity. A far-red version of this flavivirus sensor had the best signal-to-noise ratio in a fluorescent Dulbec- co’s plaque assay, leading to the construction of a multireporter platform combining the flavivirus sensor with reporter dyes for detection of chromatin condensation and cell death, enabling studies of viral plaque formation with single-cell resolution. Finally, the application of this platform enabled the study of cell-population kinetics of infection and cell death by DENV-2, ZIKV, and yellow fever virus. We anticipate that future studies of viral infection kinetics with this reporter system will enable basic investigations of virus–host interactions and facilitate future applications in antiviral drug research to manage flavivi- rus infections.es_ES
dc.description.sponsorshipInternational Centre for Genetic Engineering and Biotechnology Grant CRP/CRI18-02.es_ES
dc.language.isoenges_ES
dc.sourceJournal of Biological Chemistry, 295, 2020es_ES
dc.subjectcell deathes_ES
dc.subjectdengue virus (DENV)es_ES
dc.subjectflaviviruses_ES
dc.subjectfluorescencees_ES
dc.subjectinternal cleavagees_ES
dc.subjectNS2B-NS3 proteasees_ES
dc.subjectplaque assayes_ES
dc.subjectcell-based reporteres_ES
dc.subjectZika virus (ZIKV)es_ES
dc.subjectyellow fever virus (YFV)es_ES
dc.titleA fluorescence-activatable reporter of flavivirus NS2B–NS3 protease activity enables live imaging of infection in single cells and viral plaqueses_ES
dc.typeartículo científicoes_ES
dc.identifier.doihttps://doi.org/10.1074/jbc.RA119.011319
dc.description.procedenceUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET)es_ES
dc.description.procedenceUCR::Vicerrectoría de Docencia::Salud::Facultad de Microbiologíaes_ES


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