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

Author

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  • Matthew R. Pawlak

    (University of Minnesota)

  • Adam T. Smiley

    (University of Minnesota)

  • Maria Paz Ramirez

    (University of Minnesota)

  • Marcus D. Kelly

    (University of Minnesota)

  • Ghaidan A. Shamsan

    (University of Minnesota)

  • Sarah M. Anderson

    (University of Minnesota)

  • Branden A. Smeester

    (University of Minnesota)

  • David A. Largaespada

    (University of Minnesota)

  • David J. Odde

    (University of Minnesota)

  • Wendy R. Gordon

    (University of Minnesota)

Abstract

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.

Suggested Citation

  • Matthew R. Pawlak & Adam T. Smiley & Maria Paz Ramirez & Marcus D. Kelly & Ghaidan A. Shamsan & Sarah M. Anderson & Branden A. Smeester & David A. Largaespada & David J. Odde & Wendy R. Gordon, 2023. "RAD-TGTs: high-throughput measurement of cellular mechanotype via rupture and delivery of DNA tension probes," Nature Communications, Nature, vol. 14(1), pages 1-11, December.
  • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-38157-6
    DOI: 10.1038/s41467-023-38157-6
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    1. Benjamin L. Bangasser & Ghaidan A. Shamsan & Clarence E. Chan & Kwaku N. Opoku & Erkan Tüzel & Benjamin W. Schlichtmann & Jesse A. Kasim & Benjamin J. Fuller & Brannon R. McCullough & Steven S. Rosenf, 2017. "Shifting the optimal stiffness for cell migration," Nature Communications, Nature, vol. 8(1), pages 1-10, August.
    2. Yun Zhang & Chenghao Ge & Cheng Zhu & Khalid Salaita, 2014. "DNA-based digital tension probes reveal integrin forces during early cell adhesion," Nature Communications, Nature, vol. 5(1), pages 1-10, December.
    3. Matthew J. Paszek & Christopher C. DuFort & Olivier Rossier & Russell Bainer & Janna K. Mouw & Kamil Godula & Jason E. Hudak & Jonathon N. Lakins & Amanda C. Wijekoon & Luke Cassereau & Matthew G. Rub, 2014. "The cancer glycocalyx mechanically primes integrin-mediated growth and survival," Nature, Nature, vol. 511(7509), pages 319-325, July.
    4. Carsten Grashoff & Brenton D. Hoffman & Michael D. Brenner & Ruobo Zhou & Maddy Parsons & Michael T. Yang & Mark A. McLean & Stephen G. Sligar & Christopher S. Chen & Taekjip Ha & Martin A. Schwartz, 2010. "Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics," Nature, Nature, vol. 466(7303), pages 263-266, July.
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