Identification of proteins involved in membrane contact sites between peroxisome and ER

2014 Fiscal Year Final Research Report

• Project/Area Number: 26650060
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Cell biology
• Research Institution: Osaka University
• Principal Investigator: MAEDA Yusuke 大阪大学, 微生物病研究所, 准教授 (00294124)
• Project Period (FY): 2014-04-01 – 2015-03-31
• Keywords: オルガネラ形成・動態 / 膜接触場 / ペルオキシソーム

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.

Free Research Field

細胞生物学