Methodological approaches to studying phase separation and HIV-1 replication: Current and future perspectives.

• Bioinformatic tools are great predictors of liquid–liquid phase separation of viral proteins. • Fluorescence labelling techniques can be used to study biomolecular condensates both in vitro and in situ using confocal microscopy. • Liquid-liquid phase separation is likely to be involved in multiple steps of HIV-1 replication cycle. This article reviews tried-and-tested methodologies that have been employed in the first studies on phase separating properties of structural, RNA-binding and catalytic proteins of HIV-1. These are described here to stimulate interest for any who may want to initiate similar studies on virus-mediated liquid–liquid phase separation. Such studies serve to better understand the life cycle and pathogenesis of viruses and open the door to new therapeutics. [ABSTRACT FROM AUTHOR]