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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2015
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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. |
format | Online Article Text |
id | pubmed-4288386 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
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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