@article{d82220eb97a64f11a5481e55df5937cc,
title = "Optogenetic Reporters Delivered as mRNA Facilitate Repeatable Action Potential and Calcium Handling Assessment in Human iPSC-Derived Cardiomyocytes",
abstract = "Electrical activity and intracellular Ca2+ transients are key features of cardiomyocytes. They can be measured using organic voltage- and Ca2+sensitive dyes but their photostability and phototoxicity mean they are unsuitable for long-term measurements. Here, we investigated whether genetically encoded voltage and Ca2+ indicators (GEVIs and GECIs) delivered as modified mRNA (modRNA) into human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) would be accurate alternatives allowing measurements over long periods.These indicators were detected in hiPSC-CMs for up to 7 days after transfection and did not affect responses to proarrhythmic compounds. Furthermore, using the GEVI ASAP2f we observed action potential prolongation in long QT syndrome models, while the GECI jRCaMP1b facilitated the repeated evaluation of Ca2+ handling responses for various tyrosine kinase inhibitors.This study demonstrated that modRNAs encoding optogenetic constructs report cardiac physiology in hiPSC-CMs without toxicity or the need for stable integration, illustrating their value as alternatives to organic dyes or other gene delivery methods for expressing transgenes.",
keywords = "cardiac disease modeling, GECI, GEVI, human pluripotent stem cells, mRNA, optical analysis",
author = "Loukia Yiangou and Albert Blanch-Asensio and {de Korte}, Tessa and Miller, {Duncan C.} and {van Meer}, {Berend J.} and Mol, {Mervyn P.H.} and {van den Brink}, Lettine and Brand{\~a}o, {Karina O.} and Mummery, {Christine L.} and Davis, {Richard P.}",
note = "Funding Information: This work was supported by Starting (STEMCARDIORISK) and Proof of Concept (ACQUIRE) Grants from the European Research Council (ERC) under the European Union{\textquoteright}s Horizon 2020 Research and Innovation programme [H2020 European Research Council; grant agreements #638030 and #885469]; a VIDI fellowship from the Netherlands Organization for Scientific Research [Nederlandse Organisatie voor Wetenschappelijk Onderzoek; ILLUMINATE; #91715303]; a research grant (MONACO-SPRINT; LSHM20063) co-funded by the PPP Allowance made available by Health-Holland, Top Sector Life Sciences & Health, to stimulate public-private partnerships; and the Netherlands Organ-on-Chip Initiative, an NWO Gravitation project (024.003.001) funded by the Ministry of Education, Culture and Science of the government of the Netherlands. Panels in some figures were created using BioRender.com. Publisher Copyright: {\textcopyright} The Author(s) 2022. Published by Oxford University Press.",
year = "2022",
month = jul,
doi = "10.1093/stmcls/sxac029",
language = "English",
volume = "40",
pages = "655--668",
journal = "Stem cells",
issn = "1066-5099",
publisher = "Wiley-Blackwell",
number = "7",
}