Development of the RNAi screening system for apicobasal polarity factors
| Project/Area Number |
16H07014
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Single-year Grants |
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| Research Field |
Cell biology
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| Research Institution | The University of Tokushima |
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Principal Investigator |
HONDA Shozo 徳島大学, 大学院医歯薬学研究部(医学系), 助教 (50778206)
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| Project Period (FY) |
2016-08-26 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 上皮細胞 / 上皮極性 / RNAi / スクリーニング / ZO-1 / 免疫蛍光染色 / ゲノムワイドスクリーニング / 画像解析 / siRNA / ノックダウン |
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| Outline of Final Research Achievements |
Epithelial cells have apicobasal polarity for asymmetric cellular function to maintain homeostasis in the body. Therefore, apicobasal polarity is essential for multicellular organisms. However, the comprehensive molecular system remains to be clarified.
I developed a screening system for apicobasal polarity factors using R2/7 cells that are not capable of forming cell-cell adhesion. I found that R27 cells form apicobasal polrity and the shape of ZO-1, which is component of tight junction, was changed by knockdown of known factors in the cells. Thus, the shape of ZO-1 would be good marker of the state of the apicobasal polarity and would be applied to the screening system. Firstly, I determined appropriate immunofluorescence condition for ZO-1 in 384 well plate. Next, I established an algorithm for the detection and estimation of ZO-1 area that was ensured as acceptable assay. Lastly, I performed the screening using siRNA library and high-throughput image analysis.
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Report
(3 results)
Research Products
(1 results)
