Generation of a reporter yellow fever virus for high throughput antiviral assays.
CPER
Drug screen
HiBiT
Luciferase
Yellow fever virus
Journal
Antiviral research
ISSN: 1872-9096
Titre abrégé: Antiviral Res
Pays: Netherlands
ID NLM: 8109699
Informations de publication
Date de publication:
11 2020
11 2020
Historique:
received:
04
05
2020
revised:
16
09
2020
accepted:
19
09
2020
pubmed:
28
9
2020
medline:
16
7
2021
entrez:
27
9
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 32980446
pii: S0166-3542(20)30353-3
doi: 10.1016/j.antiviral.2020.104939
pmc: PMC7649875
pii:
doi:
Substances chimiques
Antiviral Agents
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
104939Subventions
Organisme : Medical Research Council
ID : MC_UU_12014/2
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_UU_12014/8
Pays : United Kingdom
Organisme : NIAID NIH HHS
ID : R01 AI124690
Pays : United States
Organisme : NIAID NIH HHS
ID : U19 AI057266
Pays : United States
Informations de copyright
Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.
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