Abstract
The development of arenavirus reverse genetics has provided investigators with a novel and powerful approach for the investigation of the arenavirus molecular and cell biology. The use of cell-based minigenome systems has allowed examining the cis- and trans-acting factors involved in arenavirus replication and transcription, and the identification of novel anti-arenaviral drug targets without requiring the use of live forms of arenaviruses. Likewise, it is now feasible to rescue infectious arenaviruses entirely from cloned cDNAs containing predetermined mutations in their genomes to investigate virus-host interactions and mechanisms of pathogenesis. These advances in arenavirus genetics have also facilitated screens to identify anti-arenaviral drugs and the pursuit of novel strategies to generate live-attenuated arenavirus vaccine candidates. Moreover, the generation of tri-segmented (r3) arenaviruses expressing foreign genes of interest (GOI) has opened the possibility of implementing live-attenuated arenaviruses-based vaccine vector approaches. In this chapter, we will summarize the implementation of plasmid-based reverse genetics techniques for the development of r3 arenaviruses expressing foreign GOI for their implementation as vaccine vectors.