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<p>Monitoring H<sub>2</sub>O<sub>2</sub> 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<sub>635</sub>) allows fast and sensitive chemigenetic far-red H<sub>2</sub>O<sub>2</sub> imaging while overcoming drawbacks of existing red fluorescent H<sub>2</sub>O<sub>2</sub> indicators, including oxygen dependency, high pH sensitivity, photoartifacts and intracellular aggregation. The compatibility of oROS-HT<sub>635</sub> with blue-green-shifted optical tools allows versatile optogenetic dissection of redox biology. In addition, targeted expression of oROS-HT<sub>635</sub> and multiplexed H<sub>2</sub>O<sub>2</sub> imaging enables spatially resolved imaging of H<sub>2</sub>O<sub>2</sub> targeting the plasma membrane and neighboring cells. Here we present multiplexed use cases of oROS-HT<sub>635</sub> with other green fluorescence reporters by capturing acute and real-time changes in H<sub>2</sub>O<sub>2</sub> with intracellular redox potential and Ca<sup>2+</sup> levels in response to auranofin, an inhibitor of antioxidative enzymes, via dual-color imaging. oROS-HT<sub>635</sub> enables detailed insights into intricate intracellular and intercellular H<sub>2</sub>O<sub>2</sub> dynamics, along with their interactants, through spatially resolved, far-red H<sub>2</sub>O<sub>2</sub> imaging in real time.</p><p></p>

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Monitoring in real time and far-red imaging of H2O2 dynamics with subcellular resolution

  • 期刊论文

  • 发表日期：

    2025年4月28日

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  • Justin Daho Lee,
  • Amanda Nguyen,
  • Chelsea E. Gibbs,
  • Zheyu Ruby Jin,
  • Yuxuan Wang,
  • Aida Moghadasi,
  • Sarah J. Wait,
  • Hojun Choi,
  • Kira M. Evitts,
  • Anthony Asencio,
  • Samantha B. Bremner,
  • Shani Zuniga,
  • Vedant Chavan,
  • Inez K. A. Pranoto,
  • C. Andrew Williams,
  • Annette Smith,
  • Farid Moussavi-Harami,
  • Michael Regnier,
  • David Baker,
  • Jessica E. Young,
  • David L. Mack,
  • Elizabeth Nance,
  • Patrick M. Boyle,
  • Andre Berndt

摘要

Monitoring H2O2 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-HT635) allows fast and sensitive chemigenetic far-red H2O2 imaging while overcoming drawbacks of existing red fluorescent H2O2 indicators, including oxygen dependency, high pH sensitivity, photoartifacts and intracellular aggregation. The compatibility of oROS-HT635 with blue-green-shifted optical tools allows versatile optogenetic dissection of redox biology. In addition, targeted expression of oROS-HT635 and multiplexed H2O2 imaging enables spatially resolved imaging of H2O2 targeting the plasma membrane and neighboring cells. Here we present multiplexed use cases of oROS-HT635 with other green fluorescence reporters by capturing acute and real-time changes in H2O2 with intracellular redox potential and Ca2+ levels in response to auranofin, an inhibitor of antioxidative enzymes, via dual-color imaging. oROS-HT635 enables detailed insights into intricate intracellular and intercellular H2O2 dynamics, along with their interactants, through spatially resolved, far-red H2O2 imaging in real time.