How to Measure Molecular Forces in Cells: A Guide to Evaluating Genetically-Encoded FRET-Based Tension Sensors

Abstract

The ability of cells to sense and respond to mechanical forces is central to a wide range of biological processes and plays an important role in numerous pathologies. The molecular mechanisms underlying cellular mechanotransduction, however, have remained largely elusive because suitable methods to investigate subcellular force propagation were missing. Here, we review recent advances in the development of biosensors that allow molecular force measurements. We describe the underlying principle of currently available techniques and propose a strategy to systematically evaluate new Förster resonance energy transfer (FRET)-based biosensors.

Traction force microscopy on soft elastic substrates: A guide to recent computational advances

“…1 d) [30][31][32][33][34][35][36]. Such a sensor typically consists of two molecular domains connected by a calibrated elastic linker.…”

In silico CDM model sheds light on force transmission in cell from focal adhesions to nucleus

“…Novel techniques such as genetically encoded tension sensor microscopy or intracellular tomography were developed and give an insight in intracellular forces (Cost et al, 2015;Gayrard and Borghi, 2016;Hu et al, 2003). For Contents lists available at ScienceDirect journal homepage: www.elsevier.com/locate/jbiomech www.JBiomech.com instance, FRET-based tension sensor microscopy allows intracellular tension mapping at the molecular scale identifying regions of high and low tension.…”

Pushing, pulling, and squeezing our way to understanding mechanotransduction

“…2C) [32, 33]. Recent reviews [34] discuss FRET technology in-depth and so the following discussion will highlight tools that detect forces, conformational changes, and tension-dependent changes in protein activity.…”

“…One of the most common techniques for measuring force across a protein is by engineering a variant of that protein to contain a FRET-based tension-sensing module (see [34] for more information). Such modules typically consist of a donor fluorophore separated from an acceptor fluorophore by an elastic linker domain.…”