Plasmid-Based Reverse Genetics of Influenza A Virus

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.

Keywords: Bidirectional plasmids; Influenza A virus; Plasmid-based reverse genetics; Secreted alkaline phosphatase; Virus rescue.