The 14th Annual Meeting 2008 Japan Society of Gene Therapy : Date June 12–14, 2008 Venue Sapporo Medical University School of Medicine (Auditorium) South–1, West–16, Chuo–ku, Sapporo, Hokkaido JSGT Home–page URL: http://jsgt.jp
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 618 KB
- Volume
- 11
- Category
- Article
- ISSN
- 1099-498X
- DOI
- 10.1002/jgm.1400
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✦ Synopsis
Background: Non-viral gene carriers are highly potent and promising devices for gene therapy. However, in many cases, sufficient transfection efficiency for clinical application has not been achieved and it is important to understand the intracellular processes of carriers in detail. In this study, we observed the condensation state of plasmid DNA using a CLSM after non-viral transfection and analyzed the correlation to the transgene expression. Methods: A plasmid DNA encoding Keima-Red was labeled by fluorescein and Cy3 using a method after which the DNA maintains gene expression competency even after the covalent attachment of fluorophores. After transfection using polythylenimine (polyplex) or LipofectAMINE 2000 (lipoplex) to HuH-7 cells, the fluorospectral analysis was done on the cell images by the confocal microscopy. The condensation state of plasmid DNA inside the carriers was