System Maintenance Notice

Please note that maintenance work is scheduled between 4 AM and 5 AM - CET. During this time, you may experience reduced performance. Thanks for your patience.

Publications

Journal Article Preprint Epublish 6 citations

PubMed: 38187535 DOI: 10.1101/2023.12.22.573117

Visualizing PIEZO1 Localization and Activity in hiPSC-Derived Single Cells and Organoids with HaloTag Technology.

bioRxiv

Authors: 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, Elliot E Hui, Jerome J Lacroix, Mitradas M Panicker, Srigokul Upadhyayula, Ian Parker, Medha M Pathak

Affiliations

  • Department of Physiology and Biophysics, University of California, Irvine, CA, USA. Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, CA, USA.
  • Department of Physiology and Biophysics, University of California, Irvine, CA, USA.
  • Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, CA, USA.
  • Advanced Bioimaging Center, Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
  • Center for Complex Biological Systems, University of California, Irvine, CA, USA.
  • Department of Basic Medical Sciences, Western University of Health Sciences, Pomona, CA, USA.
  • Department of Biomedical Engineering, University of California, Irvine, CA, USA.
  • Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
  • Chan Zuckerberg Biohub, San Francisco, CA, USA.
  • Department of Neurobiology and Behavior, University of California, Irvine, CA, USA.

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 Ca 2+-sensitive HaloTag ligands, achieving temporal resolution approaching that of patch clamp electrophysiology. Finally, we used lightsheet imaging of hiPSC-derived neural organoids to achieve molecular scale imaging of PIEZO1 in three-dimensional tissue organoids. Our advances offer a novel platform for studying PIEZO1 mechanotransduction in human cells and tissues, with potential for elucidating disease mechanisms and targeted therapeutic development.

MeSH

Cells, Organoids, Tissues