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Generation, Characterization, and Applications of Influenza A Reporter Viruses

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Bioluminescence

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2524))

Abstract

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.

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Acknowledgments

Influenza research in the Martinez-Sobrido laboratory is currently supported by the NIAID RO1 grants AI141607, AI145332, and HL091968 and the Department of Defense (DoD) Peer Reviewed Medical Research Program (PRMRP) grant W81XWH1710168. This work was also partly funded by the Center for Research on Influenza Pathogenesis and Transmission (CRIPT), a NIAID-funded Center of Excellence for Influenza Research and Response (CEIRR, contract # 75N93021C00014).

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Authors and Affiliations

  1. Texas Biomedical Research Institute, San Antonio, TX, USA

    Kevin Chiem & Luis Martinez-Sobrido

  2. Center for Animal Health Research, INIA-CISA/CSIC, Madrid, Spain

    Aitor Nogales

  3. Department of Internal Research, Texas Biomedical Research Institute, San Antonio, TX, USA

    Luis Martinez-Sobrido

Authors
  1. Kevin Chiem
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  2. Aitor Nogales
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  3. Luis Martinez-Sobrido
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Corresponding authors

Correspondence to Aitor Nogales or Luis Martinez-Sobrido .

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Editors and Affiliations

  1. Environmental Management Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan

    Sung-Bae Kim

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Chiem, K., Nogales, A., Martinez-Sobrido, L. (2022). Generation, Characterization, and Applications of Influenza A Reporter Viruses. In: Kim, SB. (eds) Bioluminescence. Methods in Molecular Biology, vol 2524. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2453-1_19

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  • DOI: https://doi.org/10.1007/978-1-0716-2453-1_19

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2452-4

  • Online ISBN: 978-1-0716-2453-1

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