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Article Abstract
Monitoring HO dynamics in conjunction with key biological interactants is critical for elucidating the physiological outcome of cellular redox regulation. Optogenetic hydrogen peroxide sensor with HaloTag with JF635 (oROS-HT) allows fast and sensitive chemigenetic far-red HO imaging while overcoming drawbacks of existing red fluorescent HO indicators, including oxygen dependency, high pH sensitivity, photoartifacts and intracellular aggregation. The compatibility of oROS-HT with blue-green-shifted optical tools allows versatile optogenetic dissection of redox biology. In addition, targeted expression of oROS-HT and multiplexed HO imaging enables spatially resolved imaging of HO targeting the plasma membrane and neighboring cells. Here we present multiplexed use cases of oROS-HT with other green fluorescence reporters by capturing acute and real-time changes in HO with intracellular redox potential and Ca levels in response to auranofin, an inhibitor of antioxidative enzymes, via dual-color imaging. oROS-HT enables detailed insights into intricate intracellular and intercellular HO dynamics, along with their interactants, through spatially resolved, far-red HO imaging in real time.
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| http://dx.doi.org/10.1038/s41589-025-01891-7 | DOI Listing |
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