HyPer-3: A Genetically Encoded H<sub>2</sub>O<sub>2</sub> Probe with Improved Performance for Ratiometric and Fluorescence Lifetime Imaging
Dmitry S. Bilan(Institute of Bioorganic Chemistry), Luke Pase(Karlsruhe Institute of Technology), Linda Joosen(University of Amsterdam), Andrey Yu. Gorokhovatsky(Russian Academy of Sciences), Yulia G. Ermakova(Russian Academy of Sciences), Theodorus W. J. Gadella(University of Amsterdam), Clemens Grabher(Karlsruhe Institute of Technology), Carsten Schultz(European Molecular Biology Laboratory), Sergey Lukyanov(Institute of Bioorganic Chemistry), Vsevolod V. Belousov(Institute of Bioorganic Chemistry)
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Abstract
High-performance sensors for reactive oxygen species are instrumental to monitor dynamic events in cells and organisms. Here, we present HyPer-3, a genetically encoded fluorescent indicator for intracellular H2O2 exhibiting improved performance with respect to response time and speed. HyPer-3 has an expanded dynamic range compared to HyPer and significantly faster oxidation/reduction dynamics compared to HyPer-2. We demonstrate this performance by in vivo imaging of tissue-scale H2O2 gradients in zebrafish larvae. Moreover, HyPer-3 was successfully employed for single-wavelength fluorescent lifetime imaging of H2O2 levels both in vitro and in vivo.
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