Generation, Characterization, and Applications of Influenza A Reporter Viruses.
Influenza A virus
Microneutralization assay
Recombinant viruses
Reporter genes
Reporter viruses
Reverse genetics
mCherry
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
13
7
2022
pubmed:
14
7
2022
medline:
15
7
2022
Statut:
ppublish
Résumé
Secondary experimental procedures such as immunostaining have been utilized to study wild-type influenza A viruses (IAV) but are inadequate to rapidly determine the virus in infected cells or for the high-throughput screening (HTS) of antivirals or neutralizing antibodies. Reverse genetics approaches have allowed the generation of recombinant IAV expressing bioluminescent (BL) reporters or fluorescent proteins (FPs). These approaches can easily track viral infections in cultured cells and in validated animal models of infection using in vivo imaging systems (IVIS). Here, we describe the experimental procedures to generate recombinant monomeric (m)Cherry-expressing influenza A/Puerto Rico/8/34 (PR8-mCherry) H1N1 by altering the non-structural (NS) vRNA segment and its use in mCherry-based microneutralization assays to assess antivirals and neutralizing antibodies. The experimental procedures could be used for the generation of other recombinant influenza virus types (e.g., influenza B) or IAV subtypes (e.g., H3N2) expressing mCherry or other BL reporters or FPs from the NS or other vRNA segment. These recombinant reporter-expressing viruses represent an excellent toolbox for the identification of prophylactics or therapeutics for the treatment of influenza viral infections in HTS settings as well as to study different aspects related with the biology of influenza viruses and/or its interaction with the host.
Identifiants
pubmed: 35821477
doi: 10.1007/978-1-0716-2453-1_19
pmc: PMC10285581
mid: NIHMS1901115
doi:
Substances chimiques
Antibodies, Neutralizing
0
Antiviral Agents
0
Luminescent Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
249-268Subventions
Organisme : NIAID NIH HHS
ID : R01 AI141607
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL091968
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI145332
Pays : United States
Organisme : NIAID NIH HHS
ID : 75N93021C00014
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI161175
Pays : United States
Informations de copyright
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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