Sanchez Velazquez, R. et al. (2020) Generation of a reporter yellow fever virus for high throughput antiviral assays. Antiviral Research, 183, 104939. (doi: 10.1016/j.antiviral.2020.104939) (PMID:32980446) (PMCID:PMC7649875)
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Abstract
Yellow fever virus (YFV), a member of the Flaviviridae family, is an arthropod-borne virus that can cause severe disease in humans with a lethality rate of up to 60%. Since 2017, increases in YFV activity in areas of South America and Africa have been described. Although a vaccine is available, named strain 17D (Theiler and Smith, 1937), it is contraindicated for use in the elderly, expectant mothers, immunocompromised people, among others. To this day there is no antiviral treatment against YFV to reduce the severity of viral infection. Here, we used a circular polymerase extension reaction (CPER)-based reverse genetics approach to generate a full-length reporter virus (YFVhb) by introducing a small HiBit tag in the NS1 protein. The reporter virus replicates at a similar rate to the parental YFV in HuH-7 cells. Using YFVhb, we designed a high throughput antiviral screening luciferase-based assay to identify inhibitors that target any step of the viral replication cycle. We validated our assay by using a range of inhibitors including drugs, immune sera and neutralizing single chain variable fragments (scFv). In light of the recent upsurge in YFV and a potential spread of the virus, this assay is a further tool in the development of antiviral therapy against YFV.
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Tandavanitj, Rapeepat and Sanchez Velazquez, Ricardo and Setthapramote, Dr Chayanee and De Lorenzo, Dr Giuditta and CLARK, Jordan and Varjak, Dr Margus and Patel, Professor Arvind and Kohl, Professor Alain |
| Creator Roles: | Sanchez-Velazquez, R.Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing – original draft, Writing – review and editing de Lorenzo, G.Conceptualization, Methodology Tandavanitj, R.Methodology, Formal analysis Setthapramote, C.Validation Clark, J. J.Resources, Methodology Patel, A. H.Funding acquisition, Conceptualization, Resources, Project administration, Writing – review and editing Kohl, A.Funding acquisition, Conceptualization, Resources, Writing – review and editing Varjak, M.Conceptualization, Methodology, Supervision, Writing – original draft, Writing – review and editing |
| Authors: | Sanchez Velazquez, R., de Lorenzo, G., Tandavanitj, R., Setthapramote, C., Bredenbeek, P. J., Bozzacco, L., MacDonald, M. R., Clark, J. J., Rice, C. M., Patel, A. H., Kohl, A., and Varjak, M. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research |
| Journal Name: | Antiviral Research |
| Publisher: | Elsevier |
| ISSN: | 0166-3542 |
| ISSN (Online): | 1872-9096 |
| Published Online: | 24 September 2020 |
| Copyright Holders: | Copyright © 2020 The Authors |
| First Published: | First published in Antiviral Research 183: 104939 |
| Publisher Policy: | Reproduced under a Creative Commons License |
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