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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2015-01-01
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| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fcell.2014.00074/full |
| _version_ | 1811331801218547712 |
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| author | Monika eSramkova Laura eParente Timothy eWigand Myo-Pale eAye Akiko eShitara Roberto eWeigert |
| author_facet | Monika eSramkova Laura eParente Timothy eWigand Myo-Pale eAye Akiko eShitara Roberto eWeigert |
| author_sort | Monika eSramkova |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-13T16:26:14Z |
| format | Article |
| id | doaj.art-e5c51d24079b44478cdf55ac3b3ea2b2 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-04-13T16:26:14Z |
| publishDate | 2015-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-e5c51d24079b44478cdf55ac3b3ea2b22022-12-22T02:39:44ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2015-01-01210.3389/fcell.2014.00074113502Polyethylenimine-mediated Expression of Transgenes in the Acinar Cells of Rats Salivary Glands in vivoMonika eSramkova0Laura eParente1Timothy eWigand2Myo-Pale eAye3Akiko eShitara4Roberto eWeigert5National Institute Dental And Craniofacial Research - National Institutes of HealthNational Institute Dental And Craniofacial Research - National Institutes of HealthNational Institute Dental And Craniofacial Research - National Institutes of HealthNational Institute Dental And Craniofacial Research - National Institutes of HealthNational Institute Dental And Craniofacial Research - National Institutes of HealthNational Institute Dental And Craniofacial Research - National Institutes of HealthNon 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.http://journal.frontiersin.org/Journal/10.3389/fcell.2014.00074/fullAcinar CellsAquaporin 5PolyethyleneimineSalivary Glandsintravital microscopyIn vivo Transfection |
| spellingShingle | Monika eSramkova Laura eParente Timothy eWigand Myo-Pale eAye Akiko eShitara Roberto eWeigert Polyethylenimine-mediated Expression of Transgenes in the Acinar Cells of Rats Salivary Glands in vivo Frontiers in Cell and Developmental Biology Acinar Cells Aquaporin 5 Polyethyleneimine Salivary Glands intravital microscopy In vivo Transfection |
| 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 | Acinar Cells Aquaporin 5 Polyethyleneimine Salivary Glands intravital microscopy In vivo Transfection |
| url | http://journal.frontiersin.org/Journal/10.3389/fcell.2014.00074/full |
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