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Purpose: The purpose of this study was to develop an in vivo long-term gene expression system in murine kidney using φC31 integrase. Methods: Gene expression in cultured RENCA, TCMK-1, and HEK293 cells was assessed. The long-term in vivo gene expression system in the kidney was achieved by co-transfecting 5 μg of pORF-luc/attB as a donor plasmid and 20 μg of pCMV-luc as a helper plasmid into the right kidney of mice by electroporation. Luciferase expression levels were measured to determine longevity of the expression. Results: Significantly high luciferase expression levels were observed in cultured RENCA, TCMK-1, and HEK293 cells over 1 month compared with controls (non-integrase system). The luciferase cDNA sequence was integrated at a pseudo attP site termed mpsL1. In vivo luciferase expression levels in the integrase group were sustained and significantly higher than those in the control group over 2 months. 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Long-term in vivo gene expression in mouse kidney using φC31 integrase and electroporation
http://hdl.handle.net/10069/35623
http://hdl.handle.net/10069/35623da27b555-eab8-4322-a6a9-420dcd3e2892
名前 / ファイル | ライセンス | アクション |
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JDT23_427.pdf (651.3 kB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2016-06-02 | |||||
タイトル | ||||||
タイトル | Long-term in vivo gene expression in mouse kidney using φC31 integrase and electroporation | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Electroporation | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Gene therapy | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | in vivo long-term gene expression | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Kidney | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Plasmid DNA | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Transfection | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | φC31 integrase | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者 |
Otani, Yuki
× Otani, Yuki× Kawakami, Shigeru× Mukai, Hidefumi× Fuchigami, Yuki× Yamashita, Fumiyoshi× Hashida, Mitsuru |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Background: Achieving long-term gene expression in kidney will be beneficial for gene therapy of renal and congenital diseases, genetic studies constructing animal disease models, and the functional analysis of disease-related genes. Purpose: The purpose of this study was to develop an in vivo long-term gene expression system in murine kidney using φC31 integrase. Methods: Gene expression in cultured RENCA, TCMK-1, and HEK293 cells was assessed. The long-term in vivo gene expression system in the kidney was achieved by co-transfecting 5 μg of pORF-luc/attB as a donor plasmid and 20 μg of pCMV-luc as a helper plasmid into the right kidney of mice by electroporation. Luciferase expression levels were measured to determine longevity of the expression. Results: Significantly high luciferase expression levels were observed in cultured RENCA, TCMK-1, and HEK293 cells over 1 month compared with controls (non-integrase system). The luciferase cDNA sequence was integrated at a pseudo attP site termed mpsL1. In vivo luciferase expression levels in the integrase group were sustained and significantly higher than those in the control group over 2 months. Furthermore, φC31 integrase-transfected cells had less genomic DNA damage caused by integrase expression. Discussion and conclusion: These results demonstrated that the φC31 integrase system could produce long-term (2 months) in vivo gene expression in mouse kidney. | |||||
書誌情報 |
Journal of Drug Targeting 巻 23, 号 5, p. 427-435, 発行日 2015-06-01 |
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出版者 | ||||||
出版者 | Informa Healthcare | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 1061186X | |||||
EISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 10292330 | |||||
DOI | ||||||
関連タイプ | isVersionOf | |||||
識別子タイプ | DOI | |||||
関連識別子 | 10.3109/1061186X.2014.1002788 | |||||
権利 | ||||||
権利情報 | c 2015 Informa UK Ltd. | |||||
著者版フラグ | ||||||
出版タイプ | AM | |||||
出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||
引用 | ||||||
内容記述タイプ | Other | |||||
内容記述 | Journal of Drug Targeting, 23(5), pp.427-435; 2015 |