Development of novel FP-based probes for live-cell imaging of nitric oxide dynamics

Abstract

Abstract Nitric oxide ("Equation missing") is a free radical with a wide range of biological effects, but practically impossible to visualize in single cells. Here we report the development of novel multicoloured fluorescent quenching-based "Equation missing"probes by fusing a bacteria-derived "Equation missing"-binding domain close to distinct fluorescent protein variants. These genetically encoded "Equation missing"probes, referred to as geNOps, provide a selective, specific and real-time read-out of cellular "Equation missing"dynamics and, hence, open a new era of "Equation missing"bioimaging. The combination of geNOps with a Ca 2+ sensor allowed us to visualize "Equation missing"and Ca 2+ signals simultaneously in single endothelial cells. Moreover, targeting of the "Equation missing"probes was used to detect "Equation missing"signals within mitochondria. The geNOps are useful new tools to further investigate and understand the complex patterns of "Equation missing"signalling on the single (sub)cellular level.