Real-time Monitoring of Genetically Encoded Redox Probes in Mammalian Cell Monolayers

Redox processes play an important role in cellular physiologyand pathology. A particularly powerful tool for the monitoringof cellular redox changes are genetically-encoded biosensorsbased on redox sensitive green fluorescent protein (roGFP). RoGFPscontain two cysteine residues engineered to be present on thesurface of the protein b-barrel, which are capable of forming adisulphide bond. RoGFP can be made to respond to specificredox species via the genetic fusion of appropriate redox enzymes.For example, fusion of roGFP2 to the thiol peroxidase Orp1generates an H2O2-sensitive probe.1,2