Genetic Engineering of Influenza and Other Negative-Strand RNA Viruses Containing Segmented Genomes

Abstract

Publisher Summary This chapter reviews the systems that allow the genetic manipulation of segmented negative-strand RNA viruses and summarizes how the use of reverse genetics has improved the understanding of the molecular processes of viral infection and the pathogenesis of these viruses. It discusses future perspectives of the genetic engineering of segmented negative-strand RNA viruses. Reverse genetics has had a significant impact on the understanding of replication–transcription processes and on the knowledge of the structure–function relationships of viral gene products and their biological roles. Despite their impact on influenza virus research, reverse genetics systems can be improved further. So far, systems to generate transfectant influenza viruses have been described for only six of the eight viral segments. The chapter establishes a system for the rescue of any influenza virus segment without the need for selection. Helper virus-free systems may need to be established for generating infectious virus entirely from cloned complementary DNA (cDNA), as has been done for bunyavirus; however, this may be difficult for influenza virus because of its additional gene segments. These technical limitations, however, may soon be overcome. The chapter discusses various systems used to modify influenza virus, such as influenza virus replication and reconstitution of replication-competent ribonucleo proteins. The chapter concludes with the genetic engineering of thogotoviruses and bunyaviruses.