| Project/Area Number |
18K19589
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 55:Surgery of the organs maintaining homeostasis and related fields
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| Research Institution | The University of Tokyo (2019)
Yokohama City University (2018) |
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Principal Investigator |
Sekine Keisuke 東京大学, 医科学研究所, 准教授 (00323569)
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| Project Period (FY) |
2018-06-29 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
Fiscal Year 2019: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2018: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
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| Keywords | iPS細胞 / オルガノイド / シングルセルRNAシークエンス / 肝臓 / シングルセルゲノミクス |
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| Outline of Final Research Achievements |
In this study, we constructed an efficient functional screening system for human iPSC mini-liver using CRISPR / CAS9 method. In order to efficiently carry out gene deletion, iCrispr-iPS cells in which a gene capable of inducing Cas9 expression by addition of doxycycline was introduced into the AAVS1 locus of human iPS cells by homologous recombination were established for a several iPSC lines. We performed a functional analysis using knockout iPSCs of genes that are thought to be involved in the residual undifferentiated cells in differentiated cell products, and succeeded in identifying the factors involved in the predisposed to remain the undifferentiated cells in differentiated cell products. In addition, we made several knockout iPSCs for urea cycle-related genes, which are also the cause of urea cycle abnormality, and performed functional verification.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究ではiPSCを用いて極めて効率よく遺伝子欠損細胞を作製し機能検証する手法を確立した。この技術を用いて、移植医療に用いる再生医療等製品における安全性の確保において極めて重要な未分化iPSCの残存に関わる因子を同定した。また、先天的な代謝性疾患の原因でもある尿素サイクル関連遺伝子について複数のノックアウトiPSCを作製し、尿素サイクルのそれぞれの酵素欠損によってバイパスされる代謝経路が異なる可能性が示唆された。今後、尿素サイクルに異常症に対する薬剤療法や代謝改善のための検証が可能になると期待される。以上のことから、本手法を用いることで様々な細胞における効率よい遺伝子機能の検証が期待される。
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