| Budget Amount *help |
¥181,740,000 (Direct Cost: ¥139,800,000、Indirect Cost: ¥41,940,000)
Fiscal Year 2014: ¥33,930,000 (Direct Cost: ¥26,100,000、Indirect Cost: ¥7,830,000)
Fiscal Year 2013: ¥35,620,000 (Direct Cost: ¥27,400,000、Indirect Cost: ¥8,220,000)
Fiscal Year 2012: ¥37,310,000 (Direct Cost: ¥28,700,000、Indirect Cost: ¥8,610,000)
Fiscal Year 2011: ¥36,920,000 (Direct Cost: ¥28,400,000、Indirect Cost: ¥8,520,000)
Fiscal Year 2010: ¥37,960,000 (Direct Cost: ¥29,200,000、Indirect Cost: ¥8,760,000)
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| Outline of Final Research Achievements |
FRET biosensors visualize activities of signaling molecules in living cells. However, the use was limited mostly to the cultured 2D cells. Here, we aimed at developing technologies to apply FRET biosensors in 3D culture cells and in living tissues. Due to high recombination rates, stable expression of FRET biosensors had been a difficult task. To overcome this problem, we employed the transposon-mediated gene transfer. We found either piggcBac transposase or Tol2 trasnpoase worked efficiently for the gene transfer of FRET biosensor cDNAs. In particular, we found tol2-mediated gene transfer worked efficiently to develop transgenic mice highly-expressing FRET biosensors, which we named FRET mice. By using FRET mice, we observed activation of various signaling molecules in living tissues.
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