A fluorescence-based high throughput-screening assay for the SARS-CoV RNA synthesis complex.
Antiviral Agents
/ pharmacology
Dose-Response Relationship, Drug
Drug Evaluation, Preclinical
Enzyme Activation
Fluorescent Dyes
High-Throughput Screening Assays
/ methods
Humans
Inhibitory Concentration 50
RNA, Messenger
/ genetics
RNA, Viral
RNA-Dependent RNA Polymerase
/ metabolism
Severe acute respiratory syndrome-related coronavirus
/ genetics
Severe Acute Respiratory Syndrome
/ diagnosis
Templates, Genetic
Coronavirus
Replication complex
Screening
Small molecule
Journal
Journal of virological methods
ISSN: 1879-0984
Titre abrégé: J Virol Methods
Pays: Netherlands
ID NLM: 8005839
Informations de publication
Date de publication:
02 2021
02 2021
Historique:
received:
17
07
2020
revised:
02
11
2020
accepted:
02
11
2020
pubmed:
10
11
2020
medline:
20
1
2021
entrez:
9
11
2020
Statut:
ppublish
Résumé
The Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) emergence in 2003 introduced the first serious human coronavirus pathogen to an unprepared world. To control emerging viruses, existing successful anti(retro)viral therapies can inspire antiviral strategies, as conserved viral enzymes (eg., viral proteases and RNA-dependent RNA polymerases) represent targets of choice. Since 2003, much effort has been expended in the characterization of the SARS-CoV replication/transcription machinery. Until recently, a pure and highly active preparation of SARS-CoV recombinant RNA synthesis machinery was not available, impeding target-based high throughput screening of drug candidates against this viral family. The current Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) pandemic revealed a new pathogen whose RNA synthesis machinery is highly (>96 % aa identity) homologous to SARS-CoV. This phylogenetic relatedness highlights the potential use of conserved replication enzymes to discover inhibitors against this significant pathogen, which in turn, contributes to scientific preparedness against emerging viruses. Here, we report the use of a purified and highly active SARS-CoV replication/transcription complex (RTC) to set-up a high-throughput screening of Coronavirus RNA synthesis inhibitors. The screening of a small (1520 compounds) chemical library of FDA-approved drugs demonstrates the robustness of our assay and will allow to speed-up drug discovery against the SARS-CoV-2.
Identifiants
pubmed: 33166547
pii: S0166-0934(20)30265-2
doi: 10.1016/j.jviromet.2020.114013
pmc: PMC7833800
pii:
doi:
Substances chimiques
Antiviral Agents
0
Fluorescent Dyes
0
RNA, Messenger
0
RNA, Viral
0
RNA-Dependent RNA Polymerase
EC 2.7.7.48
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
114013Informations de copyright
Copyright © 2020 Elsevier B.V. All rights reserved.
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