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Cited by 19Influenza Virus Research StudiesVirology and Viral DiseasesViral Infectious Diseases and Gene Expression in Insects

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Plasmid-Based Reverse Genetics of Influenza A Virus

Perez, Daniel R., Seibert, Brittany, Ferreri, Lucas, Lee, Chang-Won, Rajao, Daniela·2020·Methods in Molecular Biology
DOI 10.1007/978-1-0716-0346-8_4
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Abstract

Reverse genetics is the process of generating an RNA virus from a cDNA copy. Reverse genetics systems have truly transformed our ability to manipulate and study negative-strand RNA viruses. Plasmid-based reverse genetics approaches for influenza viruses provide a better understanding of virulence, transmission, mechanisms of antiviral resistance, and the development of alternative vaccines and vaccination strategies. Studying the molecular changes that allow influenza A viruses (IAVs) to transmit among animal species is important to better understand their animal health and public health risks. In this chapter, the cloning of cDNA copies of IAV's RNA segments into a reverse genetics plasmid vector, the experimental procedures for studying viral polymerase activity, and the successful generation of recombinant IAVs are described.

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Research works printed but not linked: 15

  1. King AMQ, Adams MJ, Carstens EB, Lefkowitz EJ (2012) Ninth report of the international committee on taxonomy of viruses. Elsevier Academic Press, San Diego
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  4. Sorrell EM, Wan H, Araya Y, Song H, Perez DR (2009) Minimal molecular constraints for respiratory droplet transmission of an avian-human H9N2 influenza A virus. Proc Natl Acad Sci U S A 106(18):7565–7570. https://doi.org/10.1073/pnas.0900877106 · DOI
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  6. Sutton TC, Finch C, Shao H, Angel M, Chen H, Capua I, Cattoli G, Monne I, Perez DR (2014) Airborne transmission of highly pathogenic H7N1 influenza virus in ferrets. J Virol 88(12):6623–6635. https://doi.org/10.1128/JVI.02765-13 · DOI
  7. Imai M, Watanabe T, Hatta M, Das SC, Ozawa M, Shinya K, Zhong G, Hanson A, Katsura H, Watanabe S, Li C, Kawakami E, Yamada S, Kiso M, Suzuki Y, Maher EA, Neumann G, Kawaoka Y (2012) Experimental adaptation of an influenza H5 HA confers respiratory droplet transmission to a reassortant H5 HA/H1N1 virus in ferrets. Nature 486(7403):420–428. https://doi.org/10.1038/nature10831 · DOI · Paper page
  8. Finch C, Li W, Perez DR (2015) Design of alternative live attenuated influenza virus vaccines. Curr Top Microbiol Immunol 386:205–235. https://doi.org/10.1007/82_2014_404 · DOI
  9. Hoffmann E, Neumann G, Hobom G, Webster RG, Kawaoka Y (2000) “Ambisense” approach for the generation of influenza A virus: vRNA and mRNA synthesis from one template. Virology 267(2):310–317 · DOI
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  11. Perez DR, Angel M, Gonzalez-Reiche AS, Santos J, Obadan A, Martinez-Sobrido L (2017) Plasmid-based reverse genetics of influenza A virus. Methods Mol Biol 1602:251–273. https://doi.org/10.1007/978-1-4939-6964-7_16 · DOI
  12. Nogales A, Perez DR, Santos J, Finch C, Martinez-Sobrido L (2017) Reverse genetics of influenza B viruses. Methods Mol Biol 1602:205–238. https://doi.org/10.1007/978-1-4939-6964-7_14 · DOI
  13. Te Velthuis AJW, Long JS, Barclay WS (2018) Assays to measure the activity of influenza virus polymerase. Methods Mol Biol 1836:343–374. https://doi.org/10.1007/978-1-4939-8678-1_17 · DOI
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Metadata

Authors
Perez, D. R., Seibert, B., Ferreri, L., Lee, C.-W., & Rajao, D.
Affiliations
  • University of Georgia (US)
  • The Ohio State University (US)
Corresponding author
Perez, Daniel R.
ORCID
Venue
Methods in Molecular Biology
Publisher
Springer US
Published
2020
Pages
37-59
ISSN
  • 1064-3745 (ISSN-L)
  • 1940-6029
DOI
10.1007/978-1-0716-0346-8_4
OpenAlex
W4243682365
Cited by
19
References
0
Citations by year
2026: 2 · 2025: 4 · 2024: 5 · 2022: 4 · 2021: 2 · 2020: 2
FWCI
8.52
Type
book chapter
Language
en
Open access
closed
Licence
www.springer.com/tdm (tdm)
Locations
Methods in molecular biology (publishedVersion)
Funders
National Institute of Allergy and Infectious Diseases (HHSN272201400008C), R21AI146448)
Topics
  • Health Sciences › Medicine › Epidemiology › Influenza Virus Research Studies
  • Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
Keywords
influenza A virus, cDNA cloning, negative-strand RNA viruses, recombinant virus generation, viral polymerase activity, antiviral resistance, animal-to-human transmission, influenza vaccination, viral virulence

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