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Related Experiment Video

Updated: Jan 27, 2026

Real Time Monitoring of Intracellular Bile Acid Dynamics Using a Genetically Encoded FRET-based Bile Acid Sensor
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Genetically Encoded FRET-Based Tension Sensors.

Anna-Lena Cost1,2, Samira Khalaji1,2, Carsten Grashoff1,2

  • 1Group of Molecular Mechanotransduction, Max Planck Institute of Biochemistry, Martinsried, Germany.

Current Protocols in Cell Biology
|March 14, 2019
PubMed
Summary

Genetically encoded Förster resonance energy transfer (FRET)-based tension sensors quantify molecular forces in cells. These tools are calibrated to precisely measure piconewton forces, aiding in understanding cellular mechanics.

Keywords:
FLIMFRETbiosensormechanotransductiontension sensor

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Cell Biology

Background:

  • Genetically encoded Förster resonance energy transfer (FRET)-based tension sensors are crucial for measuring piconewton-scale forces on individual molecules within living systems.
  • These biosensors exhibit high FRET efficiencies without tension, which decrease upon force application, indicating mechanical stress.

Purpose of the Study:

  • To provide a comprehensive protocol for generating and utilizing FRET-based tension sensors for specific proteins.
  • To detail methods for confirming that FRET efficiency changes accurately reflect molecular tension.
  • To enable researchers to plan, execute, and interpret FRET tension sensor experiments.

Main Methods:

  • Development of genetically encoded FRET biosensors for specific protein targets.
  • Expression of biosensors in living cells and image acquisition using live-cell fluorescence lifetime imaging microscopy (FLIM).
  • Calibration of FRET-force relationships for quantitative force measurements.
  • Detailed protocols for data analysis, statistical evaluation, and interpretation.

Main Results:

  • Demonstration of how to generate FRET-based tension sensors tailored to specific proteins of interest.
  • Establishment of controls to validate that observed FRET changes are indicative of molecular tension.
  • Methodology for relating FRET efficiency changes to calibrated mechanical forces.

Conclusions:

  • The described protocols empower researchers to effectively design and implement FRET-based tension sensor experiments.
  • This work facilitates accurate measurement and interpretation of molecular forces in cellular and organismal contexts.
  • Provides a robust framework for advancing the study of mechanobiology using FRET biosensors.