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Development of the RNAi screening system for apicobasal polarity factors

Research Project

Project/Area Number 16H07014
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeSingle-year Grants
Research Field Cell biology
Research InstitutionThe University of Tokushima

Principal Investigator

HONDA Shozo  徳島大学, 大学院医歯薬学研究部(医学系), 助教 (50778206)

Project Period (FY) 2016-08-26 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
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)
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
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.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Annual Research Report
  • Research Products

    (1 results)

All 2018

All Presentation (1 results)

  • [Presentation] 互いに接着しない上皮細胞を用いた上皮極性因子のゲノムワイドRNAiスクリーニング2018

    • Author(s)
      本田尚三、名黒功、米村重信
    • Organizer
      第3回 メカノバイオロジー学会
    • Related Report
      2017 Annual Research Report

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Published: 2016-09-02   Modified: 2019-03-29  

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