Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Direct quantification of Piezo1 activation threshold through DNA-tethered extracellular force sensing

Nature Sensors (2026)Cite this article

Subjects

Abstract

Mechanosensitive ion channels translate physical forces into biochemical signals, enabling cells to sense pressure, tension and tissue deformation. Piezo1, a trimeric mechanogated ion channel, is essential in processes such as vascular regulation and immune function, yet the force required for its activation has remained unclear because existing approaches couple applied force with membrane deformation, preventing direct calibration. Here we present a DNA-tethered extracellular force sensing platform that delivers calibrated piconewton forces directly to Piezo1 while simultaneously monitoring channel activity via a genetically encoded calcium reporter. Combining micropipette manipulation and DNA hairpin-based force calibration, we show that Piezo1 opens approximately at 15.0 pN, providing a direct quantification of its activation threshold and demonstrating that Piezo1 can be gated by tether-mediated forces independent of membrane tension, supporting a force from filament mechanism. This approach offers a generalizable strategy for precisely probing mechanotransduction pathways at the localized molecular sites and can be extended to diverse force responsive systems across biology and materials science.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to the full article PDF.

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: DNA-tethered extracellular force transduction enables direct, membrane-independent activation of Piezo1.
Fig. 2: Functional validation of engineered Piezo1–jGCaMP8m for DNA-tethered mechanical activation.
Fig. 3: DNA-tethered bead-grab reveals direct mechanical activation of Piezo1 independent of membrane deformation.
Fig. 4: Quantitative determination of the force scale for Piezo1 activation using micropipette suction and DNA hairpin calibration.

Similar content being viewed by others

Data availability

The data that support the findings of this study are present in this Article and its Supplementary Information. The full image dataset with a large size is available from the corresponding author upon request. Source data are provided with this paper.

References

  1. Wu, J., Lewis, A. H. & Grandl, J. Touch, tension, and transduction—the function and regulation of Piezoion channels. Trends Biochem. Sci. 42, 57–71 (2017).

    Article  PubMed  Google Scholar 

  2. Wang, S. et al. Endothelial cation channel PIEZO1 controls blood pressure by mediating flow-induced ATP release. J. Clin. Invest. 126, 4527–4536 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  3. Lim, X. R. et al. Endothelial Piezo1 channel mediates mechano-feedback control of brain blood flow. Nat. Commun. 15, 8686 (2024).

  4. Liu, C. S. C. et al. Piezo1 mechanosensing regulates integrin-dependent chemotactic migration in human T cells. eLife 12, RP91903 (2024).

    Article  PubMed  PubMed Central  Google Scholar 

  5. Yang, Z. et al. Nano-mechanical immunoengineering: nanoparticle elasticity reprograms tumor-associated macrophages via Piezo1. ACS Nano 18, 21221–21235 (2024).

    Article  CAS  PubMed  Google Scholar 

  6. Zarychanski, R. et al. Mutations in the mechanotransduction protein PIEZO1 are associated with hereditary xerocytosis. Blood 120, 1908–1915 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Alper, S. L. Genetic diseases of PIEZO1 and PIEZO2 dysfunction. Curr. Top. Membr. 79, 97–134 (2017).

    Article  CAS  PubMed  Google Scholar 

  8. Zhao, Q. et al. Structure and mechanogating mechanism of the Piezo1 channel. Nature 554, 487–492 (2018).

    Article  CAS  PubMed  Google Scholar 

  9. Saotome, K. et al. Structure of the mechanically activated ion channel Piezo1. Nature 554, 481–486 (2018).

    Article  CAS  PubMed  Google Scholar 

  10. Ge, J. et al. Architecture of the mammalian mechanosensitive Piezo1 channel. Nature 527, 64–69 (2015).

    Article  CAS  PubMed  Google Scholar 

  11. Kefauver, J. M., Ward, A. B. & Patapoutian, A. Discoveries in structure and physiology of mechanically activated ion channels. Nature 587, 567–576 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Lin, Y. C. et al. Force-induced conformational changes in PIEZO1. Nature 573, 230–234 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Lewis, A. H. & Grandl, J. Mechanical sensitivity of Piezo1 ion channels can be tuned by cellular membrane tension. eLife 4, e12088 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  14. Yang, S. et al. Membrane curvature governs the distribution of Piezo1 in live cells. Nat. Commun. 13, 7467 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Yang, X. et al. Structure deformation and curvature sensing of PIEZO1 in lipid membranes. Nature 604, 377–383 (2022).

    Article  CAS  PubMed  Google Scholar 

  16. Luchtefeld, I. et al. Dissecting cell membrane tension dynamics and its effect on Piezo1-mediated cellular mechanosensitivity using force-controlled nanopipettes. Nat. Methods 21, 1063–1073 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Mulhall, E. M. et al. Direct observation of the conformational states of PIEZO1. Nature 620, 1117–1125 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Wang, J. et al. Tethering Piezo channels to the actin cytoskeleton for mechanogating via the cadherin–beta-catenin mechanotransduction complex. Cell Rep. 38, 110342 (2022).

    Article  CAS  PubMed  Google Scholar 

  19. Bustamante, C., Chemla, Y. R., Forde, N. R. & Izhaky, D. Mechanical processes in biochemistry. Annu. Rev. Biochem. 73, 705–748 (2004).

    Article  CAS  PubMed  Google Scholar 

  20. Evans, E. A. & Calderwood, D. A. Forces and bond dynamics in cell adhesion. Science 316, 1148–1153 (2007).

    Article  CAS  PubMed  Google Scholar 

  21. Murthy, S. E., Dubin, A. E. & Patapoutian, A. Piezos thrive under pressure: mechanically activated ion channels in health and disease. Nat. Rev. Mol. Cell Biol. 18, 771–783 (2017).

    Article  CAS  PubMed  Google Scholar 

  22. Leckband, D. E. & de Rooij, J. Cadherin adhesion and mechanotransduction. Annu. Rev. Cell Dev. Biol. 30, 291–315 (2014).

    Article  CAS  PubMed  Google Scholar 

  23. Wang, J. et al. PIEZO1-mediated calcium signaling reinforces mechanical properties of hair follicle stem cells to promote quiescence. Sci. Adv. 11, eadt2771 (2025).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Coste, B. et al. Piezo proteins are pore-forming subunits of mechanically activated channels. Nature 483, 176–181 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Lewis, A. H. & Grandl, J. Stretch and poke stimulation for characterizing mechanically activated ion channels. Methods Enzymol. 654, 225–253 (2021).

    Article  CAS  PubMed  Google Scholar 

  26. Coste, B. et al. Piezo1 and Piezo2 are essential components of distinct mechanically activated cation channels. Science 330, 55–60 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Wang, X. et al. Mechanical stability of the cell nucleus—roles played by the cytoskeleton in nuclear deformation and strain recovery. J. Cell Sci. 131, jcs209627 (2018).

    Article  PubMed  Google Scholar 

  28. Ye, J. et al. Ultrasonic control of neural activity through activation of the mechanosensitive channel MscL. Nano Lett. 18, 4148–4155 (2018).

    Article  CAS  PubMed  Google Scholar 

  29. Peralta, F. A. et al. Optical control of PIEZO1 channels. Nat. Commun. 14, 1269 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Dumitru, A. C. et al. Probing PIEZO1 localization upon activation using high-resolution atomic force and confocal microscopy. Nano Lett. 21, 4950–4958 (2021).

    Article  CAS  PubMed  Google Scholar 

  31. Shin, W. et al. Magnetogenetics with Piezo1 mechanosensitive ion channel for CRISPR gene editing. Nano Lett. 22, 7415–7422 (2022).

    Article  CAS  PubMed  Google Scholar 

  32. Lee, J. U. et al. Non-contact long-range magnetic stimulation of mechanosensitive ion channels in freely moving animals. Nat. Mater. 20, 1029–1036 (2021).

    Article  CAS  PubMed  Google Scholar 

  33. Wu, J., Goyal, R. & Grandl, J. Localized force application reveals mechanically sensitive domains of Piezo1. Nat. Commun. 7, 12939 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Basu, A., Lagier, S., Vologodskaia, M., Fabella, B. A. & Hudspeth, A. J. Direct mechanical stimulation oftip links in hair cells through DNA tethers. eLife 5, e16041 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  35. MacHattie, L. A. & Thomas, C. A. DNA from bacteriophage lambda: molecular length and conformation. Science 144, 1142–1144 (1964).

    Article  CAS  PubMed  Google Scholar 

  36. Yaganoglu, S. et al. Highly specific and non-invasive imaging of Piezo1-dependent activity across scales using GenEPi. Nat. Commun. 14, 4352 (2023).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Gudimchuk, N. B. et al. Mechanisms of microtubule dynamics and force generation examined with computational modeling and electron cryotomography. Nat. Commun. 11, 3765 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  38. Wiegand, T. et al. Forces during cellular uptake of viruses and nanoparticles at the ventral side. Nat. Commun. 11, 32 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Huang, W., Fu, C. & Yan, J. Single-cell quantification of the mechanical stability of cell-cell adherens junction using glass micropipettes. Methods Mol. Biol. 2600, 267–280 (2023).

    Article  CAS  PubMed  Google Scholar 

  40. Huang, W., Fu, C. & Yan, J. in Mechanobiology: Methods and Protocols (ed. Zaidel-Bar, R.) 267–280 (Springer, 2023).

  41. Xiao, B. Mechanisms of mechanotransduction and physiological roles of PIEZO channels. Nat. Rev. Mol. Cell Biol. 25, 886–903 (2024).

  42. del Rio, A. et al. Stretching single talin rod molecules activates vinculin binding. Science 323, 638–641 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  43. Hoffman, B. D., Grashoff, C. & Schwartz, M. A. Dynamic molecular processes mediate cellular mechanotransduction. Nature 475, 316–323 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Elosegui-Artola, A. et al. Force triggers YAP nuclear entry by regulating transport across nuclear pores. Cell 171, 1397–1410 (2017).

    Article  CAS  PubMed  Google Scholar 

  45. Yao, M. et al. The mechanical response of talin. Nat. Commun. 7, 11966 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  46. Zhang, Y. et al. Multi-domain interaction mediated strength-building in human α-actinin dimers unveiled by direct single-molecule quantification. Nat. Commun. 15, 6151 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Zhou, T. et al. Piezo1/2 mediate mechanotransduction essential for bone formation through concerted activation of NFAT-YAP1-β-catenin. eLife 9, e52779 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Poole, K., Herget, R., Lapatsina, L., Ngo, H. D. & Lewin, G. R. Tuning Piezo ion channels to detect molecular-scale movements relevant for fine touch. Nat. Commun. 5, 3520 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

  49. Servin-Vences, M. R., Moroni, M., Lewin, G. R. & Poole, K. Direct measurement of TRPV4 and PIEZO1 activity reveals multiple mechanotransduction pathways in chondrocytes. eLife 6, e21074 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  50. Roeterink, R. M. A., Casadevall, I. S. X., Collins, D. J. & Scott, D. Force versus response: methods for activating and characterizing mechanosensitive ion channels and GPCRs. Adv. Healthc. Mater. 13, 2402167 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Andreu, I. et al. The force loading rate drives cell mechanosensing through both reinforcement and cytoskeletal softening. Nat. Commun. 12, 4229 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Hu, Y. et al. DNA-based ForceChrono probes for deciphering single-molecule force dynamics in living cells. Cell 187, 3445–3459.e15 (2024).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Research Foundation, Prime Minister’s Office, Singapore, under the NRF Investigators Program (award no. NRF-NRFI052019-0003 to X.L.) and NUS NANONASH Programme (NUHSRO/2020/002/NanoNash/LOA; award no. R143000B43114 to X.L.). J.Y. received support from the Singapore Ministry of Education Academic Research Fund Tier 2 (MOE; grant no. MOE-T2EP50123-0008), National Research Foundation, Prime Minister’s Office and the Mid-Sized Grant (grant no. NRF-MSG-2023-0001). We thank J. Grandl (Duke University) for generously providing plasmids encoding wild-type mouse Piezo1 and the bungarotoxin-binding site-inserted constructs.

Author information

Author notes

  1. These authors contributed equally: Mingyu Sui, Jingzhun Liu.

Authors and Affiliations

  1. Department of Chemistry, National University of Singapore, Singapore, Singapore

    Mingyu Sui, Yuxia Liu & Xiaogang Liu

  2. The N.1 Institute for Health, National University of Singapore, Singapore, Singapore

    Mingyu Sui, Yuxia Liu & Xiaogang Liu

  3. Joint School of National University of Singapore and Tianjin University, Tianjin University, Fuzhou, People’s Republic of China

    Mingyu Sui, Jie Yan & Xiaogang Liu

  4. Department of Physics, National University of Singapore, Singapore, Singapore

    Jingzhun Liu, Chaoyu Fu, Jie Yan & Xiaogang Liu

  5. Mechanobiology Institute, National University of Singapore, Singapore, Singapore

    Jie Yan

Authors
  1. Mingyu Sui
    View author publications

    Search author on:PubMed Google Scholar

  2. Jingzhun Liu
    View author publications

    Search author on:PubMed Google Scholar

  3. Chaoyu Fu
    View author publications

    Search author on:PubMed Google Scholar

  4. Yuxia Liu
    View author publications

    Search author on:PubMed Google Scholar

  5. Jie Yan
    View author publications

    Search author on:PubMed Google Scholar

  6. Xiaogang Liu
    View author publications

    Search author on:PubMed Google Scholar

Contributions

X.L., J.Y., M.S. and Y.L. conceived and designed the project. X.L., J.Y. and Y.L. supervised the project, providing conceptual and experimental guidance. M.S. carried our molecular cloning, electrophysiology, imaging and micropipette experiments. J.L. performed λ-DNA modifications and contributed to molecular cloning. C.F. assisted with micropipette experiments. M.S. and Y.L. drafted the original paper. X.L. and J.Y. edited the paper. All authors participated in the discussion and interpretation of the results.

Corresponding authors

Correspondence to Yuxia Liu, Jie Yan or Xiaogang Liu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

Nature Sensors thanks Isaac Li, Jisook Moon and Kate Poole for their contribution to the peer review of this work. Peer reviewer reports are available.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Effects of ionomycin treatment.

a, GFP fluorescence response to ionomycin treatment. b, Fluorescence intensity changes in HEK293T cells expressing BBS-Piezo1-jGCaMP8m upon addition of ionomycin. c, Quantification of relative fluorescence before and after ionomycin treatment (n = 24 cells from three independent tests). Statistical analysis was performed using a two-tailed paired t-test (P < 0.0001), data are presented as mean ± s.e.m.

Source data

Extended Data Fig. 2 Micropipette-enabled mechanical perturbation of GsMTx4-treated cells.

a, Representative images of HEK293T cells expressing BBS-Piezo1-jGCaMP8m during direct micropipette perturbation after GsMTx4 treatment. The dashed line indicates the position and direction of micropipette movement. b, Representative fluorescence intensity trace of GsMTx4-treated cells under micropipette perturbation. Experimental tests were repeated for three times.

Source data

Extended Data Fig. 3 Micropipette-based perturbation with DSPE-beads.

a, Bright-field images illustrating removal of untethered beads by micropipette flow. b, Representative GFP channel images showing fluorescence changes during tethered bead pulling. Experimental tests were repeated for four times.

Extended Data Fig. 4 Schematic and synthesis of the 1000 bp hairpin DNA.

The construct comprises five functional segments: a hairpin stem, a biotinylated region, a loop, a linker, and a thiol-modified end. The synthesis protocol involves four steps: (1) PCR amplification of the core hairpin sequence; (2) restriction enzyme digestion to generate cohesive ends; (3) annealing to form the hairpin structure; and (4) ligation to assemble the complete construct.

Extended Data Fig. 5 Verification of micropipette suction force using magnetic tweezers and DNA hairpin probes.

a, Force-distance calibration curve of the micropipette, with blue and green circles indicating verified force values Fclose and Fopen at corresponding pipette distances. b, Schematic of magnetic tweezer setup used to quantify hairpin closing force and opening forces. c, Bead height changes during force-jump mode (switching between two constant-force levels). d, Bead height changes during force-ramp mode (linearly increasing force). e, Schematic of micropipette suction applying lateral tension to monitor DNA hairpin opening. f, Representative trace of bead displacement versus suction pressure, showing hairpin opening under increasing force.

Source data

Supplementary information

Supplementary Information (download PDF )

Supplementary Figs. 1–12 and Tables 1 and 2.

Reporting Summary (download PDF )

Peer Review File (download PDF )

Supplementary Movie 1 (download MP4 )

Tracking of tethered GCaMP in a transfected cell.

Supplementary Movie 2 (download MP4 )

Force application via grabbing DNA-bead to activate Piezo1.

Supplementary Movie 3 (download MP4 )

Force application via remote suction on DNA-bead to activate Piezo1.

Source data

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sui, M., Liu, J., Fu, C. et al. Direct quantification of Piezo1 activation threshold through DNA-tethered extracellular force sensing. Nat. Sens. (2026). https://doi.org/10.1038/s44460-026-00060-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Version of record:

  • DOI: https://doi.org/10.1038/s44460-026-00060-0

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing