Book chapter

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

Published inInfluenza Virus. Methods in Molecular Biology, Editors Yamauchi, Yohei, p. 33-58
PublisherNew York, NY : Humana Press
Publication date2018

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.

Citation (ISO format)
ANCHISI, Stéphanie et al. Influenza A Virus Genetic Tools: From Clinical Sample to Molecular Clone. In: Influenza Virus. Methods in Molecular Biology. New York, NY : Humana Press, 2018. p. 33–58. doi: 10.1007/978-1-4939-8678-1_3
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Book chapter (Published version)

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