| Project/Area Number |
26650060
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Cell biology
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| Research Institution | Osaka University |
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Principal Investigator |
MAEDA Yusuke 大阪大学, 微生物病研究所, 准教授 (00294124)
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| Project Period (FY) |
2014-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2014: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
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| Keywords | オルガネラ形成・動態 / 膜接触場 / ペルオキシソーム |
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| Outline of Final Research Achievements |
We proved the presence of local area in mammalian cells, where peroxisome and endoplasmic reticulum membranes are closely positioned, by Proximity Ligation Assay method. Next, we tried to obtain optimal drug concentration to enrich mutant cells in which membrane contact sits (MCS) between these organelles are impaired, and the concentration was decided in MEF but not HAP1 cells. To use MEF cells, we established a new method that can directly quantify the amount of MCS in living cells by FACS using split proteins. Moreover, we used genome-wide Cas9/CRISPR gRNA library in addition to exon-trapping system. We are currently establishing new MCS mutant cells by various combination of these methods.
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