| Budget Amount *help |
¥48,230,000 (Direct Cost: ¥37,100,000、Indirect Cost: ¥11,130,000)
Fiscal Year 2015: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Fiscal Year 2014: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Fiscal Year 2013: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000)
Fiscal Year 2012: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
Fiscal Year 2011: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000)
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| Outline of Final Research Achievements |
We have developed a novel gene delivery system based on on-chip electroporation, where plasmids are directly transported into the nucleus by electrophoresis. The basic concept is examined by micro orifice fabricated on vertical PDMS wall with quantum dot and FEM simulation. The cells are cultured on micro-orifices and placed between electrodes, to which a electric pulse was applied in a DMEM medium containing GFP plasmids. Most cells showed GFP expression after pulsation, suggesting that the plasmid was directly fed into the cell nucleus. Then we applied the system to produce iPS cells. After introduction of Yamanaka factors, cells were cultured on the orifice for three weeks after electroporation, and then reprogramming of nucleus was checked. The fact that the plasmids are instantaneously transported directly into nucleus of selected cells, with precisely controllable timing, suggests that the method will find its applications in re-programming or differentiation control studies.
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