@article{0975fc3d94f549d4809a5fd8d68abcf1,
title = "Optical and electronic ion channel monitoring from native human membranes",
abstract = "Transmembrane proteins represent a major target for modulating cell activity, both in terms of therapeutics drugs and for pathogen interactions. Work on screening such therapeutics or identifying toxins has been severely limited by the lack of available methods that would give high content information on functionality (ideally multimodal) and that are suitable for high-throughput. Here, we have demonstrated a platform that is capable of multimodal (optical and electronic) screening of ligand gated ion-channel activity in human-derived membranes. The TREK-1 ion-channel was expressed within supported lipid bilayers, formed via vesicle fusion of blebs obtained from the HEK cell line overexpressing TREK-1. The resulting reconstituted native membranes were confirmed via fluorescence recovery after photobleaching to form mobile bilayers on top of films of the polymeric electroactive transducer poly(3,4-ethylenedioxythiophene) polystyrenesulfonate (PEDOT:PSS). PEDOT:PSS electrodes were then used for quantitative electrochemical impedance spectroscopy measurements of ligand-mediated TREK-1 interactions with two compounds, spadin and arachidonic acid, known to suppress and activate TREK-1 channels, respectively. PEDOT:PSS-based organic electrochemical transistors were then used for combined optical and electronic measurements of TREK-1 functionality. The technology demonstrated here is highly promising for future high-throughput screening of transmembrane protein modulators owing to the robust nature of the membrane integrated device and the highly quantitative electrical signals obtained. This is in contrast with live-cell-based electrophysiology assays (e.g., patch clamp) which compare poorly in terms of cost, usability, and compatibility with optical transduction.",
keywords = "Conducting polymer, Impedance, Ion channel, Organic electrochemical transistor, PEDOT:PSS, Supported lipid bilayer, TREK-1",
author = "Owens, {R{\'o}{\'i}s{\'i}n M.} and Susan Daniel and Pappa, {Anna Maria} and Liu, {Han Yuan} and Walther Traberg-Christensen and Quentin Thiburce and Achilleas Savva and Aimie Pavia and Alberto Salleo",
note = "Funding Information: S.D., R.O., and A.S acknowledge funding for this project, sponsored by the Defense Advanced Research Projects Agency (DARPA) Army Research Office and accomplished under Cooperative Agreement Number W911NF-18-2-0152. The views and conclusions contained in this document are those of the authors and should not be interpreted as representing the official policies, either expressed or implied, of DARPA or the Army Research Office or the U.S. Government. The U.S. Government is authorized to reproduce and distribute reprints for Government purposes notwithstanding any copyright notation herein. A.M.P. acknowledges funding from the Oppenheimer Junior Research Fellowship and the Maudslay-Butler Research Fellowship at Pembroke College, Cambridge. H.Y.L. acknowledges funding from the Fleming Fellowship at the RF Smith School of Chemical and Biomolecular Engineering at Cornell University. W.T.C. acknowledges funding from the Cambridge Commonwealth, European & International Trust atCambridge University. Part of this work was performed at the Stanford Nanofabrication Facilities (SNF) and Stanford Nano Shared Facilities (SNSF), supported by the National Science Foundation as part of the National Nanotechnology Coordinated Infrastructure under award ECCS-1542152. We also wish to acknowledge the kind gift of the HEK-TREK-1 cells from Marc Borsotto Universite de Nice Sophia Antipolis, France. Finally, we wish to acknowledge Chrysanthi-Maria Moysidou for the immunofluorescent imaging. Funding Information: S.D., R.O., and A.S acknowledge funding for this project, sponsored by the Defense Advanced Research Projects Agency (DARPA) Army Research Office and accomplished under Cooperative Agreement Number W911NF-18-2-0152. The views and conclusions contained in this document are those of the authors and should not be interpreted as representing the official policies, either expressed or implied, of DARPA or the Army Research Office or the U.S. Government. The U.S. Government is authorized to reproduce and distribute reprints for Government purposes notwithstanding any copyright notation herein. A.M.P. acknowledges funding from the Oppenheimer Junior Research Fellowship and the Maudslay-Butler Research Fellowship at Pembroke College, Cambridge. H.Y.L. acknowledges funding from the Fleming Fellowship at the RF Smith School of Chemical and Biomolecular Engineering at Cornell University. W.T.C. acknowledges funding from the Cambridge Commonwealth, European & International Trust atCambridge University. Part of this work was performed at the Stanford Nanofabrication Facilities (SNF) and Stanford Nano Shared Facilities (SNSF), supported by the National Science Foundation as part of the National Nanotechnology Coordinated Infrastructure under award ECCS-1542152. We also wish to acknowledge the kind gift of the HEK-TREK-1 cells from Marc Borsotto Universit{\'e} de Nice Sophia Antipolis, France. Finally, we wish to acknowledge Chrysanthi-Maria Moysidou for the immunofluorescent imaging. Publisher Copyright: {\textcopyright} 2020 American Chemical Society",
year = "2020",
month = oct,
day = "27",
doi = "10.1021/acsnano.0c01330",
language = "British English",
volume = "14",
pages = "12538--12545",
journal = "ACS Nano",
issn = "1936-0851",
publisher = "American Chemical Society",
number = "10",
}