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Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line

BACKGROUND: Satellite cells are residential muscle stem cells that express a paired box protein, PAX7. RESULTS: Here, we report a knock-in mouse line expressing a PAX7-enhanced yellow fluorescent protein (YFP) fusion protein that enables visualization of PAX7 protein dynamics in living satellite cel...

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Autores principales: Kitajima, Yasuo, Ono, Yusuke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108100/
https://www.ncbi.nlm.nih.gov/pubmed/30139390
http://dx.doi.org/10.1186/s13395-018-0174-x
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author Kitajima, Yasuo
Ono, Yusuke
author_facet Kitajima, Yasuo
Ono, Yusuke
author_sort Kitajima, Yasuo
collection PubMed
description BACKGROUND: Satellite cells are residential muscle stem cells that express a paired box protein, PAX7. RESULTS: Here, we report a knock-in mouse line expressing a PAX7-enhanced yellow fluorescent protein (YFP) fusion protein that enables visualization of PAX7 protein dynamics in living satellite cells through YFP fluorescence. The YFP fluorescence signals in Pax7-YFP knock-in mice clearly recapitulated the endogenous expression of PAX7 protein in satellite cells. YFP+ satellite cells were efficiently isolated from muscle tissues by fluorescence-activated cell sorting. Homozygous Pax7-YFP knock-in mice (Pax7(YFP/YFP)) were viable, grew and regenerated muscle normally, and Pax7(YFP/YFP) mouse-derived satellite cells proliferated, differentiated, and self-renewed as efficiently as those from wild-type (Pax7(+/+)) mice. CONCLUSIONS: Taken together, our Pax7-YFP mouse line is a useful tool to aid the development of stem-cell-based therapies for muscle diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13395-018-0174-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-61081002018-08-28 Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line Kitajima, Yasuo Ono, Yusuke Skelet Muscle Methodology BACKGROUND: Satellite cells are residential muscle stem cells that express a paired box protein, PAX7. RESULTS: Here, we report a knock-in mouse line expressing a PAX7-enhanced yellow fluorescent protein (YFP) fusion protein that enables visualization of PAX7 protein dynamics in living satellite cells through YFP fluorescence. The YFP fluorescence signals in Pax7-YFP knock-in mice clearly recapitulated the endogenous expression of PAX7 protein in satellite cells. YFP+ satellite cells were efficiently isolated from muscle tissues by fluorescence-activated cell sorting. Homozygous Pax7-YFP knock-in mice (Pax7(YFP/YFP)) were viable, grew and regenerated muscle normally, and Pax7(YFP/YFP) mouse-derived satellite cells proliferated, differentiated, and self-renewed as efficiently as those from wild-type (Pax7(+/+)) mice. CONCLUSIONS: Taken together, our Pax7-YFP mouse line is a useful tool to aid the development of stem-cell-based therapies for muscle diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13395-018-0174-x) contains supplementary material, which is available to authorized users. BioMed Central 2018-08-24 /pmc/articles/PMC6108100/ /pubmed/30139390 http://dx.doi.org/10.1186/s13395-018-0174-x Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
Kitajima, Yasuo
Ono, Yusuke
Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line
title Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line
title_full Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line
title_fullStr Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line
title_full_unstemmed Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line
title_short Visualization of PAX7 protein dynamics in muscle satellite cells in a YFP knock-in-mouse line
title_sort visualization of pax7 protein dynamics in muscle satellite cells in a yfp knock-in-mouse line
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108100/
https://www.ncbi.nlm.nih.gov/pubmed/30139390
http://dx.doi.org/10.1186/s13395-018-0174-x
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