RAD-TGTs: high-throughput measurement of cellular mechanotype via rupture and delivery of DNA tension probes

Mechanical forces drive critical cellular processes that are reflected in mechanical phenotypes, or mechanotypes, of cells and their microenvironment. We present here Rupture And Deliver Tension Gauge Tethers (RAD-TGTs) in which flow cytometry is used to record the mechanical history of thousands of cells exerting forces on their surroundings via their propensity to rupture immobilized DNA duplex tension probes. We demonstrate that RAD-TGTs recapitulate prior DNA tension probe studies while also yielding a gain of fluorescence in the force-generating cell that is detectable by flow cytometry. Furthermore, the rupture propensity is altered following disruption of the cytoskeleton using drugs or CRISPR-knockout of mechanosensing proteins. Importantly, RAD-TGTs can differentiate distinct mechanotypes among mixed populations of cells. We also establish oligo rupture and delivery can be measured via DNA sequencing. RAD-TGTs provide a facile and powerful assay to enable high-throughput mechanotype profiling, which could find various applications, for example, in combination with CRISPR screens and -omics analysis. Mechanical forces drive critical cellular processes, but methods to study such cellular forces are typically low-throughput. Here the authors present a method using Rupture And Deliver Tension Gauge Tethers, where flow cytometry or sequencing can be used to record the mechanical history of thousands of individual cells.

Automated summary

Mechanical forces drive critical cellular processes, and the authors present Rupture And Deliver Tension Gauge Tethers (RAD-TGTs) which can record the mechanical history of individual cells by using flow cytometry or sequencing. RAD-TGTs recapitulate prior studies on DNA tension probes and can detect fluorescence gain in the force-generating cell. They can differentiate distinct mechanotypes and have the potential for various applications in combination with CRISPR screens and -omics analysis.