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Related Experiment Video

Updated: Jul 7, 2026

Colorimetric Detection of Bacteria Using Litmus Test

Colorimetric Detection of Bacteria Using Litmus Test

Published on: September 17, 2016

Engineering redox-sensitive linkers for genetically encoded FRET-based biosensors.

Vladimir L Kolossov¹, Bryan Q Spring, Anna Sokolowski

¹Institute for Genomic Biology, Department of Animal Sciences, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA. viadimer@uiuc.edu

Experimental Biology and Medicine (Maywood, N.J.)

|January 29, 2008

Summary

Researchers developed novel genetically encoded biosensors to monitor intracellular redox environments. These biosensors, utilizing redox-sensitive linkers and Förster resonance energy transfer (FRET), accurately detect cellular oxidation states.

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Last Updated: Jul 7, 2026

Colorimetric Detection of Bacteria Using Litmus Test

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Published on: September 17, 2016

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Published on: October 1, 2016

Area of Science:

Biochemistry
Cell Biology
Biotechnology

Background:

Sensing intracellular reduction/oxidation (redox) conditions is crucial for understanding cell proliferation, differentiation, and apoptosis.
Genetically encoded biosensors offer a powerful method for real-time monitoring of the intracellular redox environment.

Purpose of the Study:

To develop novel chimeric polypeptides as redox-sensitive linkers for use with Förster resonance energy transfer (FRET) biosensors.
To create and validate genetically encoded biosensors capable of reporting on intracellular redox states.

Main Methods:

Design and synthesis of alpha-helical linkers incorporating redox-sensitive motifs, including thioredoxin-like domains and vicinal cysteines.
Construction of Förster resonance energy transfer (FRET) constructs by linking cyan and yellow fluorescent proteins with the designed redox-sensitive linkers.
Utilizing the (ratio)(A) method for robust FRET analysis and validating biosensor responsiveness in mammalian cells via flow cytometry.

Main Results:

A FRET construct incorporating the RL5 redox linker demonstrated a significant 92% increase in FRET efficiency between reduced and oxidized states.
The cyan-RL5-yellow construct successfully reported on changes in the intracellular redox environment in mammalian cells.
The developed biosensors show high sensitivity and responsiveness to redox state fluctuations.

Conclusions:

The developed redox-sensitive linkers are effective components for genetically encoded FRET biosensors.
These biosensors provide a valuable tool for studying cellular redox dynamics in various biological processes.
The study validates a novel approach for real-time intracellular redox monitoring.