A simple reverse genetics method to generate recombinant coronaviruses.
SARS-CoV-2
antivirals
in vivo experiment
reverse genetics
serology
Journal
EMBO reports
ISSN: 1469-3178
Titre abrégé: EMBO Rep
Pays: England
ID NLM: 100963049
Informations de publication
Date de publication:
04 05 2022
04 05 2022
Historique:
revised:
02
02
2022
received:
17
08
2021
accepted:
08
02
2022
pubmed:
4
3
2022
medline:
6
5
2022
entrez:
3
3
2022
Statut:
ppublish
Résumé
Engineering recombinant viruses is a pre-eminent tool for deciphering the biology of emerging viral pathogens such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the large size of coronavirus genomes renders the current reverse genetics methods challenging. Here, we describe a simple method based on "infectious subgenomic amplicons" (ISA) technology to generate recombinant infectious coronaviruses with no need for reconstruction of the complete genomic cDNA and apply this method to SARS-CoV-2 and also to the feline enteric coronavirus. In both cases we rescue wild-type viruses with biological characteristics similar to original strains. Specific mutations and fluorescent red reporter genes can be readily incorporated into the SARS-CoV-2 genome enabling the generation of a genomic variants and fluorescent reporter strains for in vivo experiments, serological diagnosis, and antiviral assays. The swiftness and simplicity of the ISA method has the potential to facilitate the advance of coronavirus reverse genetics studies, to explore the molecular biological properties of the SARS-CoV-2 variants, and to accelerate the development of effective therapeutic reagents.
Identifiants
pubmed: 35239997
doi: 10.15252/embr.202153820
pmc: PMC9066064
doi:
Substances chimiques
Antiviral Agents
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e53820Informations de copyright
© 2022 Unité des Virus Emergents. Published under the terms of the CC BY 4.0 license.
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