Abstract

Viruses are obligate intracellular pathogens that can infect all living organisms. Infections by RNA viruses are connected to many human diseases, representing a major threat for human health and a burden on economies and infrastructure. Viruses encode a limited number of proteins, and they thus rely on host resources to complete their biological cycle and this dependence represents great opportunities for antiviral intervention. The discovery of antivirals relies on the screening of large compound libraries, which also poses a technical challenge. Hence, to test the importance of cellular proteins in virus infection and to discover new antivirals, robust and efficient high throughput methods are required to measure viral fitness without using expensive reagents. Here we describe a method developed by the lab of Alfredo Castello Palomares to analyse viral gene expression in near real time using fluorescence as a proxy for infection using the CLARIOstar Plus fluorescence microplate reader with Atmospheric Control Unit (ACU). This assay has been used to understand the importance of host RNA binding proteins in viral replication to identify key proteins as potential drug targets. This method allows for the simultaneous screening of a broad range of experimental conditions, such as drug libraries and knockout cell lines in virus infection and can thus be employed for the study of host-virus interaction.