| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Outline of Final Research Achievements |
A filaggrin is one of important proteins which consist skin barrier and the mutations of this gene are reported to be essential precipitating factor of atopic dermatitis.In this study, we try to knock-out filaggrin gene (FLG) in human iPS cells and keratinocytes will be differentiated from FLG-KO hiPS cells and their parental hiPS cells. Then, finally, we will get isogenic (only FLG-KO or FLG-normal) keratinocytes. In this situation, we can evaluate the precise effect of FLG mutation (KO) to keratinocytes physiology.In order to KO FLG in hiPS cells, we decided to use engineered nucleases mediated gene targeting. First, we made FLG specific transcription activator-like effector nuclease (FLG-TALENs) , but we failed. Then, we switched to use CRISPR/Cas9 system and finally we succeeded and we got several clones of hiPS cells which possessed FLG-mutations (KO).
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