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Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo

Non viral-mediated transfection of plasmid DNA provides a fast and reliable way to express various transgenes in selected cell populations in live animals. Here, we show an improvement of a previously published method that is based on injecting plasmid DNA into the ductal system of the salivary glan...

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Autores principales: Sramkova, Monika, Parente, Laura, Wigand, Timothy, Aye, Myo-Pale', Shitara, Akiko, Weigert, Roberto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4288386/
https://www.ncbi.nlm.nih.gov/pubmed/25621283
http://dx.doi.org/10.3389/fcell.2014.00074
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author Sramkova, Monika
Parente, Laura
Wigand, Timothy
Aye, Myo-Pale'
Shitara, Akiko
Weigert, Roberto
author_facet Sramkova, Monika
Parente, Laura
Wigand, Timothy
Aye, Myo-Pale'
Shitara, Akiko
Weigert, Roberto
author_sort Sramkova, Monika
collection PubMed
description Non viral-mediated transfection of plasmid DNA provides a fast and reliable way to express various transgenes in selected cell populations in live animals. Here, we show an improvement of a previously published method that is based on injecting plasmid DNA into the ductal system of the salivary glands in live rats. Specifically, using complexes between plasmid DNA and polyethyleneimine (PEI) we show that the expression of the transgenes is directed selectively to the salivary acinar cells. PEI does not affect the ability of cells to undergo regulated exocytosis, which was one of the main drawbacks of the previous methods. Moreover PEI does not affect the proper localization and targeting of transfected proteins, as shown for the apical plasma membrane water channel aquaporin 5 (AQP5). Overall, this approach, coupled with the use of intravital microscopy, permits to conduct localization and functional studies under physiological conditions, in a rapid, reliable, and affordable fashion.
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spelling pubmed-42883862015-01-23 Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo Sramkova, Monika Parente, Laura Wigand, Timothy Aye, Myo-Pale' Shitara, Akiko Weigert, Roberto Front Cell Dev Biol Cell and Developmental Biology Non viral-mediated transfection of plasmid DNA provides a fast and reliable way to express various transgenes in selected cell populations in live animals. Here, we show an improvement of a previously published method that is based on injecting plasmid DNA into the ductal system of the salivary glands in live rats. Specifically, using complexes between plasmid DNA and polyethyleneimine (PEI) we show that the expression of the transgenes is directed selectively to the salivary acinar cells. PEI does not affect the ability of cells to undergo regulated exocytosis, which was one of the main drawbacks of the previous methods. Moreover PEI does not affect the proper localization and targeting of transfected proteins, as shown for the apical plasma membrane water channel aquaporin 5 (AQP5). Overall, this approach, coupled with the use of intravital microscopy, permits to conduct localization and functional studies under physiological conditions, in a rapid, reliable, and affordable fashion. Frontiers Media S.A. 2015-01-09 /pmc/articles/PMC4288386/ /pubmed/25621283 http://dx.doi.org/10.3389/fcell.2014.00074 Text en Copyright © 2015 Sramkova, Parente, Wigand, Aye, Shitara and Weigert. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Sramkova, Monika
Parente, Laura
Wigand, Timothy
Aye, Myo-Pale'
Shitara, Akiko
Weigert, Roberto
Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo
title Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo
title_full Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo
title_fullStr Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo
title_full_unstemmed Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo
title_short Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo
title_sort polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4288386/
https://www.ncbi.nlm.nih.gov/pubmed/25621283
http://dx.doi.org/10.3389/fcell.2014.00074
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