splitFAST: a split fluorescent reporter with rapid and reversible complementation

First publication of splitFAST

Dr. Arnaud Gautier and Dr. Alison Tebo from ENS (Paris, France) report in Nature Commun. the development of splitFAST, a fluorescence complementation system for the visualization of transient protein-protein interactions in living cells. Engineered from the fluorogenic reporter FAST (Fluorescence-Activating and absorption-Shifting Tag), splitFAST displays rapid and reversible complementation, allowing the real-time visualization of both the formation and the dissociation of a protein assembly. The paper discloses various applications of splitFAST: interaction between a membrane protein and a cytosolic protein, interactions from the MAPK signaling pathway, real-time monitoring of the MEK1-ERK2 interaction, real-time monitoring of transient Ca-dependent interactions, apoptosis biosensor.

Beyond FRET and BiFC

Conventional imaging techniques for protein-protein interactions encompass F?ster Resonance Energy Transfer (FRET) and bimolecular fluorescence complementation (BiFC), the latter being found easy to implement, straightforward to interpret, and less sensitive to the relative levels of the two interacting proteins. Now, in contrast to BiFC, splitFAST complementation was shown fully reversible and disassembly rapid, which allows not only the real-time monitoring of protein complex assembly but also the real-time monitoring of protein complex disassembly. This unprecedented behavior opens exciting prospects to study the role and function of protein-protein interactions in various cellular processes and dissect complex interaction networks.

splitFAST works with the classical fluorogens of The Twinkle Factory, reflecting the versatility of this breakthrough fluorescent reporting system.

#splitFAST #FRET #BiFC