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Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors
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A color-tailored fluorogenic sensor for hydrogen peroxide.

Ekaterina S Potekhina1,2,3, Dina I Bass2,3, Daria Ezeriņa4,5,6

  • 1Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, Moscow, Russia.

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|October 16, 2025
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Summary
This summary is machine-generated.

Scientists developed HyPerFLEX, a novel sensor for precise hydrogen peroxide (H₂O₂) monitoring in living cells. This advanced tool allows detailed imaging of cellular redox signaling and oxidative stress dynamics.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Imaging

Background:

  • Hydrogen peroxide (H₂O₂) is a crucial signaling molecule regulating cellular metabolism and involved in oxidative stress.
  • Existing sensors have limitations in detecting low H₂O₂ levels or imaging in specific cellular compartments.

Purpose of the Study:

  • To develop a novel sensor, HyPerFLEX, for high-precision, oxygen-independent monitoring of H₂O₂ dynamics in living cells.
  • To enable tunable spectral imaging of H₂O₂ across different cellular compartments, including under hypoxic conditions.

Main Methods:

  • HyPerFLEX combines the Neisseria meningitidis OxyR domain with the Y-FAST fluorogenic protein.
  • Oxidation of OxyR by H₂O₂ triggers oxygen-independent fluorescence.
  • Tunable excitation spectra allow for green to far-red emission.

Main Results:

  • HyPerFLEX demonstrates superior sensitivity for detecting ultralow H₂O₂ concentrations compared to HyPer7.
  • The sensor successfully imaged H₂O₂ dynamics during early glucose-stimulated insulin production.
  • HyPerFLEX enabled H₂O₂ measurement in the oxidizing environment of the endoplasmic reticulum lumen.

Conclusions:

  • HyPerFLEX is a powerful and versatile tool for advanced H₂O₂ imaging in living cells.
  • Its capabilities extend to studying oxidative stress and cellular signaling with high precision.
  • The sensor's tunable spectra and sensitivity offer new possibilities for multicompartment and low-concentration H₂O₂ analysis.