Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice
We have previously developed an efficient and safe transfection method for the kidney in mice: renal suction-mediated transfection. In this study, we verified the detailed characteristics of transgene expression and plasmid DNA (pDNA) in mice to develop therapeutic strategies and application to gene...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2017-01-01
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| Series: | Drug Delivery |
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| Online Access: | http://dx.doi.org/10.1080/10717544.2017.1333171 |
| _version_ | 1819177175465590784 |
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| author | Natsuko Oyama Yuki Fuchigami Shintaro Fumoto Megumu Sato Masayori Hagimori Kazunori Shimizu Shigeru Kawakami |
| author_facet | Natsuko Oyama Yuki Fuchigami Shintaro Fumoto Megumu Sato Masayori Hagimori Kazunori Shimizu Shigeru Kawakami |
| author_sort | Natsuko Oyama |
| collection | DOAJ |
| description | We have previously developed an efficient and safe transfection method for the kidney in mice: renal suction-mediated transfection. In this study, we verified the detailed characteristics of transgene expression and plasmid DNA (pDNA) in mice to develop therapeutic strategies and application to gene function analysis in the kidney. After naked pDNA was administered intravenously, the right kidney was immediately suctioned by a tissue suction device. We examined the spatial distribution of transgene expression and pDNA in the suctioned kidney using tissue clearing by CUBIC, ClearT2, and Scale SQ reagents. Spatial distribution analysis showed that pDNA was transfected into extravascular cells and sufficiently delivered to the deep renal cortex. In addition, we revealed that transgene expression occurred mainly in peritubular fibroblasts of the suctioned kidney by tissue clearing and immunohistochemistry. Next, we confirmed the periods of pDNA uptake and activation of transcription factors nuclear factor-κB and activator protein 1 by luciferase assays. Moreover, the use of a pCpG-free plasmid enabled sustained transgene expression in the suctioned kidney. In conclusion, analyses of the spatial distribution and immunostaining of the section suggest that pDNA and transgene expression occurs mainly in peritubular fibroblasts of the suctioned kidney. In addition, we clarified some factors for efficient and/or sustained transgene expression in the suctioned kidney. |
| first_indexed | 2024-12-22T21:22:29Z |
| format | Article |
| id | doaj.art-ed8e09f4657846a5839c8e630b5049dc |
| institution | Directory Open Access Journal |
| issn | 1071-7544 1521-0464 |
| language | English |
| last_indexed | 2024-12-22T21:22:29Z |
| publishDate | 2017-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Drug Delivery |
| spelling | doaj.art-ed8e09f4657846a5839c8e630b5049dc2022-12-21T18:12:09ZengTaylor & Francis GroupDrug Delivery1071-75441521-04642017-01-0124190691710.1080/10717544.2017.13331711333171Characterization of transgene expression and pDNA distribution of the suctioned kidney in miceNatsuko Oyama0Yuki Fuchigami1Shintaro Fumoto2Megumu Sato3Masayori Hagimori4Kazunori Shimizu5Shigeru Kawakami6Nagasaki UniversityNagasaki UniversityNagasaki UniversityNagasaki UniversityNagasaki UniversityNagoya UniversityNagasaki UniversityWe have previously developed an efficient and safe transfection method for the kidney in mice: renal suction-mediated transfection. In this study, we verified the detailed characteristics of transgene expression and plasmid DNA (pDNA) in mice to develop therapeutic strategies and application to gene function analysis in the kidney. After naked pDNA was administered intravenously, the right kidney was immediately suctioned by a tissue suction device. We examined the spatial distribution of transgene expression and pDNA in the suctioned kidney using tissue clearing by CUBIC, ClearT2, and Scale SQ reagents. Spatial distribution analysis showed that pDNA was transfected into extravascular cells and sufficiently delivered to the deep renal cortex. In addition, we revealed that transgene expression occurred mainly in peritubular fibroblasts of the suctioned kidney by tissue clearing and immunohistochemistry. Next, we confirmed the periods of pDNA uptake and activation of transcription factors nuclear factor-κB and activator protein 1 by luciferase assays. Moreover, the use of a pCpG-free plasmid enabled sustained transgene expression in the suctioned kidney. In conclusion, analyses of the spatial distribution and immunostaining of the section suggest that pDNA and transgene expression occurs mainly in peritubular fibroblasts of the suctioned kidney. In addition, we clarified some factors for efficient and/or sustained transgene expression in the suctioned kidney.http://dx.doi.org/10.1080/10717544.2017.1333171gene deliverykidneynaked pdnaspatial distributionfibroblast |
| spellingShingle | Natsuko Oyama Yuki Fuchigami Shintaro Fumoto Megumu Sato Masayori Hagimori Kazunori Shimizu Shigeru Kawakami Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice Drug Delivery gene delivery kidney naked pdna spatial distribution fibroblast |
| title | Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice |
| title_full | Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice |
| title_fullStr | Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice |
| title_full_unstemmed | Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice |
| title_short | Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice |
| title_sort | characterization of transgene expression and pdna distribution of the suctioned kidney in mice |
| topic | gene delivery kidney naked pdna spatial distribution fibroblast |
| url | http://dx.doi.org/10.1080/10717544.2017.1333171 |
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