Abstract

The present invention relates to a new optical tool for detecting and quantifying pyruvate in samples, in tissues and in cellular and subcellular compartments, with high spatial and temporal resolution, this is a Forster Resonance Energy Transfer (FRET)-based pyruvate sensor comprising a bacterial PdhR transcription factor between any suitable donor and acceptor fluorescent proteins moieties. The invention also relates to methods of use of this novel optical tool for the quantification of the activity of pyruvate transporters, for the quantification of the rates of cellular pyruvate production and consumption, and for the direct quantification of the rate of mitochondrial pyruvate consumption in intact cells.