| Project/Area Number |
17550159
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemistry related to living body
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| Research Institution | Osaka City University |
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Principal Investigator |
NAGASAKI Takeshi Osaka City University, Graduate School of Engineering, Associate Professor, 大学院工学研究科, 助教授 (30237507)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2005: ¥2,800,000 (Direct Cost: ¥2,800,000)
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| Keywords | Biomaterials / Nano-biotechnology / Gene / Biotechnology / Gene delivery / Non-viral vector / Transfection / Cell biology / 非ウイルスベクター / 生体機能利用 / 細胞内動態 / エンドサイトーシス |
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| Research Abstract |
The effective gene delivery system is an essential element for successful achievement of not only the gene therapy but also exhaustive analyses of genes at post-genome era. The non-viral vector has to act as a molecular machinery like a virus infecting into host cells and amplifying efficiently in order to overcome extracellular and intracellular barriers and has to lead a smooth mRNA transcription at nuclei as a final destination. In this project, stimulus responsive gene delivery systems in which some structural factors and/or physiological properties are regulated responding extracellular signals such as light are developed as a new generation of non-viral vectors. In order to enhance the nuclear import of exogenous genes, novel plasmid DNA/importin-β conjugates, which consist of a biotinylated plasmid DNA and a recombinant streptavidin-fused importin-β, were prepared. The microinjection of plasmid DNA/importin-β conjugates into the cytoplasm of NIH3T3 cells resulted in the nuclear
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localization of conjugates and the higher expression efficiency, compared to intact plasmid DNA alone. These results indicate that plasmid DNA/importin-β conjugates would be an important tool to enhance the nuclear localization of exogenous DNA in non-viral gene delivery system. Moreover, the enhancement of internalization efficiency by dual ligands is an important factor for improving transfection efficiency. In addition, the effect of dual ligands depends on cell species; receptor-mediated and efficient internalization may be related to this enhanced transfection efficiency. Biodegradable polymeric vectors were obtained from chitosan and polylysines.
Obtained results on development of artificial multi-component vectors aimed at providing solutions to membrane crossing, endosomal escape and navigation through the nuclear pore. The combination of these nature-inspired and chemically originated approaches is bringing us continually closer to the concept of constructing an artificial virus capable of delivering viable nucleic acid-based pharmaceuticals to defined cells as molecular machines that can work even in vivo. Less
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