| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
|
|---|
| Outline of Final Research Achievements |
With our established systems of obtaining iKCs from hiPSCs and new technology of programmable nucleases, especially CRISPR/Cas9 system, we tried to clarify the precise effects of filaggrin gene (FLG) mutations in keratinocytes. A guide RNA that targeted appropriate site of human FLG was designed by web-based tool and cloning into the backbone vector of CRIPSR/Cas9 (hFLG-CRISPR/Cas9). We transfected hFLG-CRISPR/Cas9 into hiPSCs and obtained the several clones of hiPSCs which possessed random mutations in FLG. Then, original hiPSC and FLG-mutated hiPSCs were differentiated into epidermal keratinocytes using our established protocols and we obtained the normal iKCs and FLG-mutated iKCs. Under this condition, we can compare the phenotypes of normal and FLG-mutaed iKCs of the same genetic background.
Thus, the results obtained from this system should be "true" meanings of FLG mutation in keratinocytes and should be important information for the understanding of AD pathogenesis.
|
