Generation of a reporter yellow fever virus for high throughput antiviral assays

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 > Institute of Infection Immunity and Inflammation
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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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
656491Basis of the host range and tissue tropism for hepatitis C virusArvind PatelMedical Research Council (MRC)MC_UU_12014/2MVLS III - CENTRE FOR VIRUS RESEARCH
656551Arbovirus interactions with arthropod hostsAlain KohlMedical Research Council (MRC)MC_UU_12014/8MVLS III - CENTRE FOR VIRUS RESEARCH