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Nat Commun. 2016 Feb 4;7:10623. doi: 10.1038/ncomms10623.

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

Author information

1
Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21/III, 8010 Graz, Austria.
2
Institute of Biophysics, Center of Physiological Medicine, Medical University of Graz, Harrachgasse 21/IV, 8010 Graz, Austria.
3
Institute of Physiological Chemistry, Center of Physiological Medicine, Medical University of Graz, Harrachgasse 21/II, 8010 Graz, Austria.
4
Nanomedical Research Laboratory, Department of Chemistry and Biochemistry, Ohio University, 350 West State Street, Athens, Ohio 45701, USA.

Abstract

Nitric oxide () 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 probes by fusing a bacteria-derived -binding domain close to distinct fluorescent protein variants. These genetically encoded probes, referred to as geNOps, provide a selective, specific and real-time read-out of cellular dynamics and, hence, open a new era of bioimaging. The combination of geNOps with a Ca(2+) sensor allowed us to visualize and Ca(2+) signals simultaneously in single endothelial cells. Moreover, targeting of the probes was used to detect signals within mitochondria. The geNOps are useful new tools to further investigate and understand the complex patterns of signalling on the single (sub)cellular level.

PMID:
26842907
PMCID:
PMC4743004
DOI:
10.1038/ncomms10623
[Indexed for MEDLINE]
Free PMC Article

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