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Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells
Published on: March 16, 2017
Real-Time Imaging of Nitric Oxide Signals in Individual Cells Using geNOps
Emrah Eroglu1, Helmut Bischof1, Suphachai Charoensin1
1Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria.
Abstract:
Nitric oxide (NO•) is a versatile signaling molecule which regulates fundamental cellular processes in all domains of life. However, due to the radical nature of NO• it has a very short half-life that makes it challenging to trace its formation, diffusion, and degradation on the level of individual cells. Very recently, we expanded the family of genetically encoded sensors by introducing a novel class of single fluorescent protein-based NO• probes-the geNOps. Once expressed in cells of interest, geNOps selectively respond to NO• by fluorescence quench, which enables real-time monitoring of cellular NO• signals. Here, we describe detailed methods suitable for imaging of NO• signals in mammalian cells. This novel approach may facilitate a broad range of studies to (re)investigate the complex NO• biochemistry in living cells.
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