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Quantification of virus-infected cells using RNA FISH-Flow

We present a protocol to detect cells that have been infected by RNA viruses. The method, RNA fluorescence in situ hybridization flow cytometry (RNA FISH-Flow), uses 48 fluorescently labeled DNA probes that hybridize in tandem to viral RNA. RNA FISH-Flow probes can be synthesized to match any RNA vi...

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Detalles Bibliográficos
Autores principales: Warren, Cody J., Barbachano-Guerrero, Arturo, Huey, Devra, Yang, Qing, Worden-Sapper, Emma R., Kuhn, Jens H., Sawyer, Sara L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10209735/
https://www.ncbi.nlm.nih.gov/pubmed/37209094
http://dx.doi.org/10.1016/j.xpro.2023.102291
Descripción
Sumario:We present a protocol to detect cells that have been infected by RNA viruses. The method, RNA fluorescence in situ hybridization flow cytometry (RNA FISH-Flow), uses 48 fluorescently labeled DNA probes that hybridize in tandem to viral RNA. RNA FISH-Flow probes can be synthesized to match any RNA virus genome, in either sense or anti-sense, enabling detection of genomes or replication intermediates within cells. Flow cytometry enables high-throughput analysis of infection dynamics within a population at the single cell level. For complete details on the use and execution of this protocol, please refer to Warren et al. (2022).(1)