Genetic biosensors for imaging nitric oxide in single cells

• Published in: Free radical biology & medicine Vol. 128; pp. 50 - 58
• Main Authors: Eroglu, Emrah, Charoensin, Suphachai, Bischof, Helmut, Ramadani, Jeta, Gottschalk, Benjamin, Depaoli, Maria R., Waldeck-Weiermair, Markus, Graier, Wolfgang F., Malli, Roland
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20-11-2018
• Subjects: Biosensing Techniques - methods; Fluorescence microscopy; Fluorescent Dyes - chemistry; FRET; Genetically encoded fluorescent probes; Humans; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Microscopy, Fluorescence; Nitric oxide; Nitric Oxide - analysis; Nitric Oxide - metabolism; Signal Transduction; Single cell imaging; FRET; Single cell imaging; Genetically encoded fluorescent probes; Nitric oxide; Fluorescence microscopy
• ISSN: 0891-5849; 1873-4596
• DOI: 10.1016/j.freeradbiomed.2018.01.027

Over the last decades a broad collection of sophisticated fluorescent protein-based probes was engineered with the aim to specifically monitor nitric oxide (NO), one of the most important signaling molecules in biology. Here we report and discuss the characteristics and fields of applications of currently available genetically encoded fluorescent sensors for the detection of NO and its metabolites in different cell types.
Because of its radical nature and short half-life, real-time imaging of NO on the level of single cells is challenging. Herein we review state-of-the-art genetically encoded fluorescent sensors for NO and its byproducts such as peroxynitrite, nitrite and nitrate. Such probes enable the real-time visualization of NO signals directly or indirectly on the level of single cells and cellular organelles and, hence, extend our understanding of the spatiotemporal dynamics of NO formation, diffusion and degradation. Here, we discuss the significance of NO detection in individual cells and on subcellular level with genetic biosensors. Currently available genetically encoded fluorescent probes for NO and nitrogen species are critically discussed in order to provide insights in the functionality and applicability of these promising tools. As an outlook we provide ideas for novel approaches for the design and application of improved NO probes and fluorescence imaging protocols.
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•Genetically encoded fluorescent probes are indispensable tools in modern cell research.•Specific fluorescent biosensors allow detection of nitric oxide (NO) in individual cells.•Direct and indirect genetically encoded NO probes are available.•Genetically encoded NO probes will be improved in future.