A major challenge in combating significant human pathogens is the inherent variation of viruses that can evolve to evade the host immune response and antiviral treatment strategies. Our research proposes an interdisciplinary approach to identify novel antiviral targets by interrogating the host cell biology of infection by influenza A virus (IAV), human immunodeficiency virus (HIV) and hepatitis C virus (HCV) using high-throughput RNA interference (RNAi) technologies. More specifically, we propose to understand the dependency of these enveloped viruses on the host endoplasmic reticulum (ER) machinery for virus replication and propagation and how this increased virus burden activates host-ER stress, including the unfolded protein response (UPR). The UPR is implicated in inflammatory signaling and we will further investigate whether these viruses can modulate host immune responses via this pathway. In summary, we have proposed a hypothesis-driven RNAi screen to generate an overview of common host-cell factors exploited by diverse viruses during infection as well as extensive biochemical and functional validation of any sites of conserved virus-protein interactions. Therefore, this work aims to identify novel candidates for further translational research and development as improved, broad-spectrum antiviral treatments to combat infection and associated pathogenesis.