| Project/Area Number |
18K06953
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 49010:Pathological biochemistry-related
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
Kurata Morito 東京医科歯科大学, 大学院医歯学総合研究科, 講師 (40451926)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | がん遺伝子 / CRISPR screening / RAS / NRAS / 癌遺伝子 / CRISPR library / CRISPR activation / 分子標的薬 / スクリーニング / CRISPR |
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| Outline of Final Research Achievements |
Mutations in RAS family are observed in 6-96% of cancer. However, the role of the magnitude of RAS activation in proliferation and cell cycle progression is not fully understood. To define the role of oncogenic NRAS dose in malignant behavior, we used a genetically engineered THP-1 cell line where the expression of NRAS-G12V is under the control of a doxycycline-responsive element. We established modified cell lines, named B11. To identify whole downstream of NRAS signals, CRISPR libraries were introduced into B11 and removing doxycycline to collect clones that can proliferate without NRAS signaling. As a result, 21 from the activation library. NRAS activation with Dox can induce the mRNA expression for DOHH and TAF6 in B11. DOHH and TAF6 are newly identified as a downstream of NRAS signaling
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| Academic Significance and Societal Importance of the Research Achievements |
RASシグナルの下流として考えられているRAF-MEK-ERK-pathwayを阻害することで、一部の癌細胞では治療効果が期待できる。しかしながら、すべてのRAS変異癌細胞を制御する包括的な治療法の確立には至っていない。網羅的な解析を行えるCRISPR library によるスクリーニングを使用することで、NRASのシグナル経路に関わる新たな遺伝子を同定した。今後、更に詳細な検討が必要とされるものの、RAS経路の全貌を明らかにできる手法の確立に成功したと考えられる。
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