Development of a FRET-based recombinant tension sensor to visualize cell-material interactions

Area of Science:

• Biophysics
• Cell Biology
• Biomaterials Science

Background:

• Förster/fluorescence resonance energy transfer (FRET)-based molecular tension sensors have elucidated intracellular mechanotransduction.
• The applicability of recombinant tension sensors for detecting extracellular forces remains largely unexplored.

Purpose of the Study:

• To develop and validate a recombinant FRET-based tension sensor (rFRET-TS) for measuring forces in the extracellular environment.
• To investigate the sensor's ability to visualize cell-material mechanical interactions.

Main Methods:

• Development of a recombinant FRET-based tension sensor (rFRET-TS).
• Immobilization of the rFRET-TS onto a glass surface with an RGDS peptide.
• Seeding of fibroblasts onto the sensor-coated surface.
• Time-lapse FRET imaging to monitor sensor dynamics and cellular forces.

Main Results:

• The rFRET-TS successfully detected forces generated by fibroblasts interacting with the material surface.
• Intra-molecular FRET was dominant, indicating sensor tension.
• Real-time visualization of forces related to focal adhesion formation and actin cytoskeletal reorganization was achieved.

Conclusions:

• The developed rFRET-TS is effective for real-time visualization of cell-material mechanical interactions.
• This sensor holds promise for understanding cell-scaffold interactions and guiding cell behavior control.