Protocol to evaluate the antiviral effect of FDA-approved drugs against dengue virus in Huh7 cells and AG129 mice

• Published in: STAR protocols 2024-06, Vol.5 (2), p.102992, Article 102992
• Main Authors: Palacios-Rápalo, Selvin Noé, Hernández-Castillo, Jonathan, Cordero-Rivera, Carlos Daniel, Benítez-Vega, Magda Lizbeth, De Jesús-González, Luis Adrián, Reyes-Ruiz, José Manuel, Farfan-Morales, Carlos Noe, Osuna-Ramos, Juan Fidel, Gonzalez-Gonzalez, Arely M., Cruz, Raymundo, del Ángel, Rosa María
• Format: Article
• Language: English
• Subjects: Animals; Antiviral Agents - pharmacology; Cell Biology; Cell Line; Dengue - drug therapy; Dengue - virology; Dengue Virus - drug effects; Flow Cytometry; Health Sciences; Humans; Mice; Microbiology; Microscopy; Model Organisms; Protocol; United States; United States Food and Drug Administration
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2024.102992

Finding an effective therapy against diseases caused by flaviviruses remains a challenge. Here, we present a protocol to test Food and Drug Administration-approved drugs that inhibit host nuclear protein import, promoting a reduction of dengue infection. We describe steps for analyzing the drug effect on nuclear import inhibition of cellular and viral proteins by confocal microscopy or western blotting. We then describe procedures for measuring the antiviral drug effects on virus-infected cells by flow cytometry and testing drug efficacy in dengue-infected AG129 mice by survival assays. For complete details on the use and execution of this protocol, please refer to Palacios-Rápalo et al.1 [Display omitted] •Protocol for measuring the antiviral effect of FDA-approved drugs on DENV infection•Procedures for measuring the FDA-approved drugs effect on DENV proteins nuclear import•Measurement of effect of nuclear import inhibitors on DENV proteins localization in vivo Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Finding an effective therapy against diseases caused by flaviviruses remains a challenge. Here, we present a protocol to test Food and Drug Administration-approved drugs that inhibit host nuclear protein import, promoting a reduction of dengue infection. We describe steps for analyzing the drug effect on nuclear import inhibition of cellular and viral proteins by confocal microscopy or western blotting. We then describe procedures for measuring the antiviral drug effects on virus-infected cells by flow cytometry and testing drug efficacy in dengue-infected AG129 mice by survival assays.