ESCRT is a machinery that enables the severing of membrane necks in a variety of cellular processes. Among the ESCRT family, ESCRT-III is thought to be the driving force for the membrane fission reaction by polymerizing into helical structures. Until recently, the main technique used to label these complexes, and other proteins, in live cells was by tagging them with fluorescent (Fl) proteins. The main problem with such approach is that Fl-proteins are larger than most ESCRT-III subunits and consistently tagging ESCRT-III with Fl-proteins interferes with proper function of these proteins. We therefore, set to apply a more direct approach for labeling these proteins in live cells.
