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KAKEN_24601016seika.pdf
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Download
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:138.0 KB
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:Apr 11, 2016 |
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Total downloads since Apr 11, 2016 : 271
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放射線障害評価に有用なレポーター細胞の開発と幹細胞技術を用いた評価系の確立
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ホウシャセン ショウガイ ヒョウカ ニ ユウヨウナ レポーター サイボウ ノ カイハツ ト カンサイボウ ギジュツ オ モチイタ ヒョウカケイ ノ カクリツ
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Hoshasen shogai hyoka ni yuyona repota saibo no kaihatsu to kansaibo gijutsu o mochiita hyokakei no kakuritsu
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Establishment of the reporter stem cell lines capable of evaluating the cell influence by radiation.
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今井, 貴雄
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イマイ, タカオ
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Imai, Takao
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慶應義塾大学・医学部・助教
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Research team head
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科研費研究者番号 : 10383712
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河瀬, 聡
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カワセ, サトシ
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Kawae, Satoshi
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慶應義塾大学・医学部・特任助教
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Research team member
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科研費研究者番号 : 80468496
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西本, 祥仁
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ニシモト, ヨシノリ
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Nishimoto, Yoshinori
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慶應義塾大学・医学部・特任助教
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Research team member
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科研費研究者番号 : 30398622
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2015
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科学研究費補助金研究成果報告書
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2014
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細胞の染色体に存在する遺伝子に対して障害性を有する放射線(ガンマ線, X線など)について, その障害を評価し得るレポーター細胞の創出を本研究は目的とした。DNA組み換え技術により, p53遺伝子座を含むBAC DNA(RP11-89D11)上にオワンクラゲ由来蛍光蛋白質とホタル由来発光蛋白質の融合遺伝子(dVenusLuc2)を挿入置換したBACレポーター遺伝子の作成を完了した。また, 近年発達してきたゲノム改変技術(CRISPR-CAS9法)により, H9 ES細胞由来神経幹細胞株の内在性p53遺伝子座にレポーター遺伝子dVenusLuc2が適切に挿入されている細胞を選別し, 株化を達成した。
The present study was intended to generate the reporter stem cells capable of evaluating the cell influence by radiation (gamma ray, X-ray), which cause double strand break of chromosomal DNA. By DNA recombination technology, the fusion reporter gene (dVenusLuc2: Aequorea victoria-derived fluorescence protein and firefly-derived light-emitting protein) was inserted to the p53 locus on BAC DNA RP11-89D11. Thus, the generation of the p53 BAC reporter gene was completed. In addition, by using recently developed genome modification technique CRISPR-CAS9 method, the reporter gene dVenusLuc2 is properly inserted into the endogenous p53 locus of H9 ES cell-derived neural stem cell lines. We achieve the establishment of neural stem cell line that contain the fluorescent and luminescent reporter gene on their p53 genomic locus.
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研究種目 : 基盤研究(C)
研究期間 : 2012～2014
課題番号 : 24601016
研究分野 : 分子生物学, 幹細胞生物学
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