A multifunctional mitochondrial genetic probe to understand mitochondrial biology.

The Stagi lab has developed a set of transfectable/transducable probes which allow the study of mitochondria in biochemical, proteomic and physiological applications. Briefly, the probe is genetically encoded and incorporated in the mitochondria. The set is composed of: Mitochondrial markers in different colours recruited to outer or inner membranes with localized biotinylation systems for spin-free isolation. Coupled to this are probes with Ca2+, pH and membrane potential sensors which allow live imaging, and FACS sorting so that mitochondria with specific characteristics can be isolated. This allows coupled live imaging and proteomic applications in cell lines. The project will involve optimising techniques to isolate and study mitochondria, using the state of the art of imaging, biochemistry enzymology and physiology, to develop robust assays for specific mito-proteomics/metabolomics studies. We will compile proteomic assays of mitochondria isolated with this new innovative biochemical system. If the testing and completion of this set of tools will be successful is certainly have a large application in study the physiology of mitochondria directly in the organelle without contamination.