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In vivo Electroporation of Skeletal Muscle Fibers in Mice

In vitro models are essential for investigating the molecular, biochemical, and cell-biological aspects of skeletal muscle. Still, models that utilize cell lines or embryonic cells do not fully recapitulate mature muscle fibers in vivo. Protein function is best studied in mature differentiated tissu...

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Detalles Bibliográficos
Autores principales: Foltz, Steven J., Hartzell, Criss H., Choo, Hyojung J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bio-Protocol 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10339231/
https://www.ncbi.nlm.nih.gov/pubmed/37456334
http://dx.doi.org/10.21769/BioProtoc.4759
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author Foltz, Steven J.
Hartzell, Criss H.
Choo, Hyojung J.
author_facet Foltz, Steven J.
Hartzell, Criss H.
Choo, Hyojung J.
author_sort Foltz, Steven J.
collection PubMed
description In vitro models are essential for investigating the molecular, biochemical, and cell-biological aspects of skeletal muscle. Still, models that utilize cell lines or embryonic cells do not fully recapitulate mature muscle fibers in vivo. Protein function is best studied in mature differentiated tissue, where biological context is maintained, but this is often difficult when reliable detection reagents, such as antibodies, are not commercially available. Exogenous expression of tagged proteins in vivo solves some of these problems, but this approach can be technically challenging because either a mouse must be engineered for each protein of interest or viral vectors are required for adequate levels of expression. While viral vectors can infect target cells following local administration, they carry the risk of genome integration that may interfere with downstream analyses. Plasmids are another accessible expression system, but they require ancillary means of cell penetration; electroporation is a simple physical method for this purpose that requires minimal training or specialized equipment. Here, we describe a method for in vivo plasmid expression in a foot muscle following electroporation.
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spelling pubmed-103392312023-07-14 In vivo Electroporation of Skeletal Muscle Fibers in Mice Foltz, Steven J. Hartzell, Criss H. Choo, Hyojung J. Bio Protoc Methods Article In vitro models are essential for investigating the molecular, biochemical, and cell-biological aspects of skeletal muscle. Still, models that utilize cell lines or embryonic cells do not fully recapitulate mature muscle fibers in vivo. Protein function is best studied in mature differentiated tissue, where biological context is maintained, but this is often difficult when reliable detection reagents, such as antibodies, are not commercially available. Exogenous expression of tagged proteins in vivo solves some of these problems, but this approach can be technically challenging because either a mouse must be engineered for each protein of interest or viral vectors are required for adequate levels of expression. While viral vectors can infect target cells following local administration, they carry the risk of genome integration that may interfere with downstream analyses. Plasmids are another accessible expression system, but they require ancillary means of cell penetration; electroporation is a simple physical method for this purpose that requires minimal training or specialized equipment. Here, we describe a method for in vivo plasmid expression in a foot muscle following electroporation. Bio-Protocol 2023-07-05 /pmc/articles/PMC10339231/ /pubmed/37456334 http://dx.doi.org/10.21769/BioProtoc.4759 Text en ©Copyright : © 2023 The Authors; This is an open access article under the CC BY-NC license https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the CC BY-NC license (https://creativecommons.org/licenses/by-nc/4.0/).
spellingShingle Methods Article
Foltz, Steven J.
Hartzell, Criss H.
Choo, Hyojung J.
In vivo Electroporation of Skeletal Muscle Fibers in Mice
title In vivo Electroporation of Skeletal Muscle Fibers in Mice
title_full In vivo Electroporation of Skeletal Muscle Fibers in Mice
title_fullStr In vivo Electroporation of Skeletal Muscle Fibers in Mice
title_full_unstemmed In vivo Electroporation of Skeletal Muscle Fibers in Mice
title_short In vivo Electroporation of Skeletal Muscle Fibers in Mice
title_sort in vivo electroporation of skeletal muscle fibers in mice
topic Methods Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10339231/
https://www.ncbi.nlm.nih.gov/pubmed/37456334
http://dx.doi.org/10.21769/BioProtoc.4759
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