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A Photoprotein in Mouse Embryonic Stem Cells Measures Ca(2+) Mobilization in Cells and in Animals
Exogenous expression of pharmacological targets in transformed cell lines has been the traditional platform for high throughput screening of small molecules. However, exogenous expression in these cells is limited by aberrant dosage, or its toxicity, the potential lack of interaction partners, and a...
Autores principales: | , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2811732/ https://www.ncbi.nlm.nih.gov/pubmed/20111708 http://dx.doi.org/10.1371/journal.pone.0008882 |
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author | Cainarca, Silvia Fenu, Simone Ferri, Cinzia Nucci, Cinzia Arioli, Patrizia Menegon, Andrea Piemonti, Lorenzo Lohmer, Stefan Wrabetz, Lawrence Corazza, Sabrina |
author_facet | Cainarca, Silvia Fenu, Simone Ferri, Cinzia Nucci, Cinzia Arioli, Patrizia Menegon, Andrea Piemonti, Lorenzo Lohmer, Stefan Wrabetz, Lawrence Corazza, Sabrina |
author_sort | Cainarca, Silvia |
collection | PubMed |
description | Exogenous expression of pharmacological targets in transformed cell lines has been the traditional platform for high throughput screening of small molecules. However, exogenous expression in these cells is limited by aberrant dosage, or its toxicity, the potential lack of interaction partners, and alterations to physiology due to transformation itself. Instead, primary cells or cells differentiated from precursors are more physiological, but less amenable to exogenous expression of reporter systems. To overcome this challenge, we stably expressed c-Photina, a Ca(2+)-sensitive photoprotein, driven by a ubiquitous promoter in a mouse embryonic stem (mES) cell line. The same embryonic stem cell line was also used to generate a transgenic mouse that expresses c-Photina in most tissues. We show here that these cells and mice provide an efficient source of primary cells, cells differentiated from mES cells, including cardiomyocytes, neurons, astrocytes, macrophages, endothelial cells, pancreatic islet cells, stably and robustly expressing c-Photina, and may be exploited for miniaturized high throughput screening. Moreover, we provide evidence that the transgenic mice may be suitable for ex-vivo bioimaging studies in both cells and tissues. |
format | Text |
id | pubmed-2811732 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-28117322010-01-29 A Photoprotein in Mouse Embryonic Stem Cells Measures Ca(2+) Mobilization in Cells and in Animals Cainarca, Silvia Fenu, Simone Ferri, Cinzia Nucci, Cinzia Arioli, Patrizia Menegon, Andrea Piemonti, Lorenzo Lohmer, Stefan Wrabetz, Lawrence Corazza, Sabrina PLoS One Research Article Exogenous expression of pharmacological targets in transformed cell lines has been the traditional platform for high throughput screening of small molecules. However, exogenous expression in these cells is limited by aberrant dosage, or its toxicity, the potential lack of interaction partners, and alterations to physiology due to transformation itself. Instead, primary cells or cells differentiated from precursors are more physiological, but less amenable to exogenous expression of reporter systems. To overcome this challenge, we stably expressed c-Photina, a Ca(2+)-sensitive photoprotein, driven by a ubiquitous promoter in a mouse embryonic stem (mES) cell line. The same embryonic stem cell line was also used to generate a transgenic mouse that expresses c-Photina in most tissues. We show here that these cells and mice provide an efficient source of primary cells, cells differentiated from mES cells, including cardiomyocytes, neurons, astrocytes, macrophages, endothelial cells, pancreatic islet cells, stably and robustly expressing c-Photina, and may be exploited for miniaturized high throughput screening. Moreover, we provide evidence that the transgenic mice may be suitable for ex-vivo bioimaging studies in both cells and tissues. Public Library of Science 2010-01-27 /pmc/articles/PMC2811732/ /pubmed/20111708 http://dx.doi.org/10.1371/journal.pone.0008882 Text en Cainarca et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Cainarca, Silvia Fenu, Simone Ferri, Cinzia Nucci, Cinzia Arioli, Patrizia Menegon, Andrea Piemonti, Lorenzo Lohmer, Stefan Wrabetz, Lawrence Corazza, Sabrina A Photoprotein in Mouse Embryonic Stem Cells Measures Ca(2+) Mobilization in Cells and in Animals |
title | A Photoprotein in Mouse Embryonic Stem Cells Measures Ca(2+) Mobilization in Cells and in Animals |
title_full | A Photoprotein in Mouse Embryonic Stem Cells Measures Ca(2+) Mobilization in Cells and in Animals |
title_fullStr | A Photoprotein in Mouse Embryonic Stem Cells Measures Ca(2+) Mobilization in Cells and in Animals |
title_full_unstemmed | A Photoprotein in Mouse Embryonic Stem Cells Measures Ca(2+) Mobilization in Cells and in Animals |
title_short | A Photoprotein in Mouse Embryonic Stem Cells Measures Ca(2+) Mobilization in Cells and in Animals |
title_sort | photoprotein in mouse embryonic stem cells measures ca(2+) mobilization in cells and in animals |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2811732/ https://www.ncbi.nlm.nih.gov/pubmed/20111708 http://dx.doi.org/10.1371/journal.pone.0008882 |
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