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Influenza Virus pp 33–58Cite as

Influenza A Virus Genetic Tools: From Clinical Sample to Molecular Clone

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1836))

Abstract

Implementation of reverse genetics for influenza A virus, that is, the DNA-based generation of infectious viral particles in cell culture, opened new avenues to investigate the function of viral proteins and their interplay with host factors on a molecular level. This powerful technique allows the introduction, depletion, or manipulation of any given sequence in the viral genome, as long as it gives rise to replicating virus progeny. Reverse genetics can be used to generate targeted reassortant viruses by mixing segments of different viral strains, thus providing insight into phenotypes of potentially pandemic viruses arising from natural reassortment. It was further instrumental for the development of novel vaccine strategies, allowing rapid and targeted exchange of viral surface antigens on a well-replicating genetic backbone of cell culture-adapted or cold-adapted/attenuated viral strains. Establishment of reverse genetics and rescue of molecular clones of influenza A virus have been extensively described before. Here we give a detailed stand-alone protocol encompassing clinical sampling of influenza A virus specimens and subsequent plasmid-based genetics to rescue, manipulate, and confirm a fully infectious molecular clone. This protocol is based on the combined techniques and experience of a number of influenza laboratories, which are credited and referenced whenever appropriate.

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Author information

Author notes

  1. Stéphanie Anchisi, Ana Rita Goncalves, and Béryl Mazel-Sanchez contributed equally to this work.

Authors and Affiliations

  1. Faculty of Medicine, CMU, Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, Switzerland

    Stéphanie Anchisi, Béryl Mazel-Sanchez & Mirco Schmolke

  2. Laboratory of Virology, Infectious Diseases Service, University Hospitals of Geneva, Geneva, Switzerland

    Ana Rita Gonçalves & Samuel Cordey

Authors
  1. Stéphanie Anchisi
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  2. Ana Rita Gonçalves
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  3. Béryl Mazel-Sanchez
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  4. Samuel Cordey
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  5. Mirco Schmolke
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Corresponding authors

Correspondence to Samuel Cordey or Mirco Schmolke .

Editor information

Editors and Affiliations

  1. School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK

    Yohei Yamauchi

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Anchisi, S., Gonçalves, A.R., Mazel-Sanchez, B., Cordey, S., Schmolke, M. (2018). Influenza A Virus Genetic Tools: From Clinical Sample to Molecular Clone. In: Yamauchi, Y. (eds) Influenza Virus. Methods in Molecular Biology, vol 1836. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8678-1_3

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  • DOI: https://doi.org/10.1007/978-1-4939-8678-1_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8677-4

  • Online ISBN: 978-1-4939-8678-1

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