Measuring cellular force using DNA-based tension probes: from ensemble to single-molecule studies

Area of Science:

• Cellular mechanobiology
• Molecular biophysics
• Biotechnology

Background:

• Cellular mechanical forces are vital for processes like adhesion, migration, differentiation, and immune activation.
• Understanding mechanotransduction is key to deciphering these cellular behaviors.

Purpose of the Study:

• To describe two advanced DNA-based tension probes for studying mechanotransduction in living cells.
• To provide a detailed protocol for the application of these probes.

Main Methods:

• Development and application of reversible shearing DNA-based tension probes (RSDTPs) for ensemble force measurements (4–60 pN).
• Utilizing ForceChrono probes for dynamic, single-molecule force measurements, capturing magnitude, duration, and loading rate.
• Detailed procedures for probe synthesis, purification, cell experiments, imaging, and computational data analysis.

Main Results:

• RSDTPs enable reversible, dynamic quantification of cellular forces without ligand depletion.
• ForceChrono probes offer insights into the temporal dynamics of single-molecule force transmission.
• The protocol facilitates detailed analysis of cellular mechanobiology, including integrin mechanobiology and cell adhesion.

Conclusions:

• These DNA-based tension probes are powerful tools for quantitative analysis of cellular mechanical forces.
• The described protocol enables comprehensive study of mechanotransduction with applications in various cell biology fields.
• The probes and methods are accessible to researchers with relevant backgrounds and can be implemented within a few days.