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dc.creatorArias Arias, Jorge Luis
dc.creatorMora Rodríguez, Rodrigo Antonio
dc.date.accessioned2022-07-20T13:25:50Z
dc.date.available2022-07-20T13:25:50Z
dc.date.issued2021-03-05
dc.identifier.citationhttps://bio-protocol.org/default.aspxes_ES
dc.identifier.issn2331-8325
dc.identifier.urihttps://hdl.handle.net/10669/86987
dc.description.abstractThe genus Flavivirus within the family Flaviviridae includes many viral species of medical importance, such as yellow fever virus (YFV), Zika virus (ZIKV), and dengue virus (DENV), among others. Presently, the identification of flavivirus-infected cells is based on either the immunolabeling of viral proteins, the application of recombinant reporter replicons and viral genomes, or the use of cellbased molecular reporters of the flaviviral protease NS2B-NS3 activity. Among the latter, our flavivirusactivatable GFP and mNeptune reporters contain a quenching peptide (QP) joined to the fluorescent protein by a linker consisting of a cleavage site for the flavivirus NS2B-NS3 proteases (AAQRRGRIG). When the viral protease cleaves the linker, the quenching peptide is removed, and the fluorescent protein adopts a conformation promoting fluorescence. Here we provide a detailed protocol for the generation, selection and implementation of stable BHK-21 cells expressing our flavivirus geneticallyencoded molecular reporters, suitable to monitor the viral infection by live-cell imaging. We also describe the image analysis procedures and provide the required software pipelines. Our reporter cells allow the implementation of single-cell infection kinetics as well as plaque assays for both reference and native strains of flaviviruses by live-cell imaging.es_ES
dc.description.sponsorshipVicerrectoría de Investigación, Universidad de Costa Rica, proyecto 803-B9-505 International Centre for Genetic Engineering and Biotechnology Grant CRP/CRI18-02.es_ES
dc.language.isoenges_ES
dc.sourceBioProtocol 11(05), 2021es_ES
dc.subjectFlaviviruses_ES
dc.subjectFluorescencees_ES
dc.subjectNS2B-NS3es_ES
dc.subjectProteasees_ES
dc.subjectLive-cell imaginges_ES
dc.subjectReporter cellses_ES
dc.subjectPlaque assayes_ES
dc.subjectImage analysises_ES
dc.titleGeneration and implementation of reporter BHK-21 cells for live imaging of flavivirus infectiones_ES
dc.typeartículo científicoes_ES
dc.identifier.doi10.21769/BioProtoc.3942
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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