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Long-term in vivo gene expression in mouse kidney using φC31 integrase and electroporation


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Title: Long-term in vivo gene expression in mouse kidney using φC31 integrase and electroporation
Authors: Otani, Yuki / Kawakami, Shigeru / Mukai, Hidefumi / Fuchigami, Yuki / Yamashita, Fumiyoshi / Hashida, Mitsuru
Issue Date: 1-Jun-2015
Publisher: Informa Healthcare
Citation: Journal of Drug Targeting, 23(5), pp.427-435; 2015
Abstract: 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.
Keywords: Electroporation / Gene therapy / in vivo long-term gene expression / Kidney / Plasmid DNA / Transfection / φC31 integrase
URI: http://hdl.handle.net/10069/35623
ISSN: 1061186X
DOI: 10.3109/1061186X.2014.1002788
Rights: © 2015 Informa UK Ltd.
Type: Journal Article
Text Version: author
Appears in Collections:Articles in academic journal

Citable URI : http://hdl.handle.net/10069/35623

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