| Project/Area Number |
14571126
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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 |
General surgery
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MIYATA Tetsuro The University of Tokyo, Faculty of Medicine, Associate Professor, 医学部附属病院, 助教授 (70190791)
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| Co-Investigator(Kenkyū-buntansha) |
HAMADA Hirofumi Sapporo Medical University, Dept. of Molecular Medicine, Professor, 分子医学研究部門, 教授 (00189614)
SAKUMA Ichiro The University of Tokyo, Graduate school and faculty of engineering, Professor, 大学院・新領域創成科学研究科, 教授 (50178597)
SHIGEMATSU Hiroshi The University of Tokyo, Faculty of Medicine, Associate Professor, 医学部附属病院, 助教授 (40134556)
SHIGEMATSU Kunihiro The University of Tokyo, Faculty of Medicine, Research Associate, 医学部附属病院, 助手 (20215966)
KOYAMA Hiroyuki The University of Tokyo, Faculty of Medicine, Associate Professor, 医学部附属病院, 助教授 (10241994)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | electroporation / plasmid DNA / gene transfer |
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| Research Abstract |
We developed a nonviral method for arterial gene transfer by in vivo electroporation with plasmid DNA. Electroporation is the physical process of including nanometreaized transient pores in a cell membrane by the application of short duration, high intensity electric field pulses to cells or tissues. The most important parameters for effective electroporation are: the stimulation voltage, the pulse duration, the electric current, plasmid concentration, and the shape of an electrode.
Result: Electroporation with 30 voltage and pulse duration of 20 msec resulted in a marked increase of gene expression in vessel wall. The concentration of 400μ g/ml plasmid DNA is most optimal. The Bite of the vessel gene-transferred by in vivo electroporation is focused around the edge of the electrode. The same findings are demonstrated by the simulation model of vessel wall and the experiment of optical mapping. We compare with gene expression in vessel wall between with the monoplate electrode, which is one 5^*1mm plate, and the multiplate electrode, which is two plates in parallel. The gene expression using muitiplate electrode is superior to the one of monoplate electrode.
In summary, we developed a method for gene transfer using electroporation, an approach which proved to the safe and more efficient than that using naked DNA. And then it will be possible that the site or the mount of gene expression is handled in the future.
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