| Project/Area Number |
18K15288
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 50020:Tumor diagnostics and therapeutics-related
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| Research Institution | Nagoya University (2019)
Saitama Medical University (2018) |
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Principal Investigator |
Nitta Hirohisa 名古屋大学, 生命農学研究科, 研究員 (60808690)
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| Project Period (FY) |
2018-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2019: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2018: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | 遺伝子修復治療 / 遺伝子治療 / ゲノム編集 / 造血幹細胞 / CIRCLE-seq / iCaspase9 / マーモセット / オフターゲット / IL2RG / 免疫不全 |
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| Outline of Final Research Achievements |
In order to carry out safe therapeutic genome editing, an attempt was made to establish a highly efficient and safe technique using human hematopoietic stem cells for therapeutic genome editing using X-linked severe combined immunodeficiency marmoset. As a result, it was confirmed that the homologous recombination efficiency reached the target value and the undifferentiated state was maintained. The off-target sites were confirmed by using the CIRCLE-seq method, but the sites at which the gene function was lost could not be detected. At the same time, I attempted to develop a technique for removing cells in which the donor sequence was inserted at a site different from the target. Suicide gene iCaspase9 incorporated into the donor sequence reduced the rate of cells with non-homologous recombination.
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| Academic Significance and Societal Importance of the Research Achievements |
ゲノム編集技術を用いて正しい位置に正常遺伝子（ドナー配列）を導入して、変異遺伝子を正常遺伝子に置き換える遺伝子修復治療が可能となった。常染色体優性遺伝病の治療は遺伝子修復治療のみ治療効果が期待できるが、CRISPRが標的以外の類似配列を切断するオフターゲット変異、ドナー配列の標的領域以外への挿入などの安全性の問題点がある。本研究では、長期的な効率性と安全性の評価のためにマーモセットを用いることを目標として遺伝子修復治療技術を確立させた。同時に、ドナー配列に新しい自殺遺伝子を組み込むことにより安全な遺伝子修復治療技術の開発を行なった。これらの成果は遺伝子修復治療技術の発展に結びつく。
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