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Visualizing PIEZO1 localization and activity in hiPSC-derived single cells and organoids with HaloTag technology

Author

Listed:
  • Gabriella A. Bertaccini

    (University of California
    University of California)

  • Ignasi Casanellas

    (University of California
    University of California)

  • Elizabeth L. Evans

    (University of California
    University of California)

  • Jamison L. Nourse

    (University of California
    University of California)

  • George D. Dickinson

    (University of California)

  • Gaoxiang Liu

    (University of California)

  • Sayan Seal

    (University of California)

  • Alan T. Ly

    (University of California
    University of California)

  • Jesse R. Holt

    (University of California
    University of California
    University of California)

  • Tharaka D. Wijerathne

    (Western University of Health Sciences)

  • Shijun Yan

    (University of California)

  • Elliot E. Hui

    (University of California)

  • Jerome J. Lacroix

    (Western University of Health Sciences)

  • Mitradas M. Panicker

    (University of California
    University of California)

  • Srigokul Upadhyayula

    (University of California
    Lawrence Berkeley National Laboratory
    Chan Zuckerberg Biohub)

  • Ian Parker

    (University of California
    University of California)

  • Medha M. Pathak

    (University of California
    University of California
    University of California
    University of California)

Abstract

PIEZO1 is critical to numerous physiological processes, transducing diverse mechanical stimuli into electrical and chemical signals. Recent studies underscore the importance of visualizing endogenous PIEZO1 activity and localization to understand its functional roles. To enable physiologically and clinically relevant studies on human PIEZO1, we genetically engineered human induced pluripotent stem cells (hiPSCs) to express a HaloTag fused to endogenous PIEZO1. Combined with advanced imaging, our chemogenetic platform allows precise visualization of PIEZO1 localization dynamics in various cell types. Furthermore, the PIEZO1-HaloTag hiPSC technology facilitates the non-invasive monitoring of channel activity across diverse cell types using Ca2+-sensitive HaloTag ligands, achieving temporal resolution approaching that of patch clamp electrophysiology. Finally, we use lightsheet microscopy on hiPSC-derived neural organoids to achieve molecular scale imaging of PIEZO1 in three-dimensional tissue. Our advances establish a platform for studying PIEZO1 mechanotransduction in human systems, with potential for elucidating disease mechanisms and targeted drug screening.

Suggested Citation

  • Gabriella A. Bertaccini & Ignasi Casanellas & Elizabeth L. Evans & Jamison L. Nourse & George D. Dickinson & Gaoxiang Liu & Sayan Seal & Alan T. Ly & Jesse R. Holt & Tharaka D. Wijerathne & Shijun Yan, 2025. "Visualizing PIEZO1 localization and activity in hiPSC-derived single cells and organoids with HaloTag technology," Nature Communications, Nature, vol. 16(1), pages 1-18, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-59150-1
    DOI: 10.1038/s41467-025-59150-1
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