Abstract |
Induced Pluripotent Stem Cells (iPSCs) can be differentiated into epithelial organoids that recapitulate the relevant context for CFTR and enable testing of therapies targeting Cystic Fibrosis (CF)-causing mutant proteins. However, to date, CF-iPSC-derived organoids have only been used to study pharmacological modulation of mutant CFTR channel activity and not the activity of other disease-relevant membrane protein constituents. In the current work, we describe a high-throughput, fluorescence-based assay of CFTR channel activity in iPSC-derived intestinal organoids and describe how this method can be adapted to study other apical membrane proteins. Specifically, we show how this assay can be employed to study CFTR and ENaC channels and an electrogenic acid transporter in the same iPSC-derived intestinal tissue. This phenotypic platform promises to expand CF therapy discovery to include strategies that target multiple determinants of epithelial fluid transport.
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Authors | Sunny Xia, Zoltán Bozóky, Michelle Di Paola, Onofrio Laselva, Saumel Ahmadi, Jia Xin Jiang, Amy L Pitstick, Chong Jiang, Daniela Rotin, Christopher N Mayhew, Nicola L Jones, Christine E Bear |
Journal | Cells
(Cells)
Vol. 10
Issue 12
(12 04 2021)
ISSN: 2073-4409 [Electronic] Switzerland |
PMID | 34943927
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Amino Acid Transport Systems
- Epithelial Sodium Channels
- SLC6A14 protein, human
- cystic fibrosis transmembrane conductance regulator delta F508
- Cystic Fibrosis Transmembrane Conductance Regulator
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Topics |
- Amino Acid Transport Systems
(metabolism)
- Animals
- Cell Differentiation
- Cystic Fibrosis Transmembrane Conductance Regulator
(metabolism)
- Dogs
- Epithelial Sodium Channels
(metabolism)
- Gene Editing
- Humans
- Induced Pluripotent Stem Cells
(metabolism)
- Intestines
(metabolism)
- Madin Darby Canine Kidney Cells
- Organoids
(metabolism)
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