Establishment of the reporter stem cell lines capable of evaluating the cell influence by radiation.

• Project/Area Number: 24601016
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: 医学物理学・放射線技術学
• Research Institution: Keio University
• Principal Investigator: IMAI TAKAO 慶應義塾大学, 医学部, 助教 (10383712)
• Co-Investigator(Renkei-kenkyūsha): KAWAE Satoshi 慶應義塾大学, 医学部, 特任助教 (80468496) ; NISHIMOTO Yoshinori 慶應義塾大学, 医学部, 特任助教 (30398622)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 幹細胞 / 神経幹細胞 / ゲノム編集 / p53 / 放射線 / レポーター遺伝子 / DNA傷害 / 蛍光蛋白質

Outline of Final Research Achievements

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.

Research Products

• [Journal Article] The long non-coding RNA nuclear-enriched abundant transcript 1_2 induces paraspeckle formation in the motor neuron during the early phase of amyotrophic lateral sclerosis (2013)
  • Author(s): Nishimoto, Y., Nakagawa, S., Hirose, T., Okano, H. J., Takao, M., Shibata, S., Suyama, S., Kuwako, K., Imai, T., Murayama, S., Suzuki, N., and Okano, H
  • Journal Title: Mol. Brain Volume: 6 Issue: 1 Pages: 31-31
  • DOI: 10.1186/1756-6606-6-31
  • Peer Reviewed
• [Journal Article] Musashi-1 post-transcriptionally enhances phosphotyrosine-binding domain- containing m-Numb protein expression in regenerating gastric mucosa. (2013)
  • Author(s): Tetsufumi Takahashi
  • Journal Title: PLoS One Volume: 8 Issue: 1 Pages: 1-13
  • DOI: 10.1371/journal.pone.0053540
  • Peer Reviewed
• [Journal Article] Cellular toxicity induced by the 26-kDa fragment and amyotrophic lateral sclerosis-associated mutant forms of TAR DNA-binding protein 43 in human embryonic stem cell-derived motor neurons. (2013)
  • Author(s): Nishimoto Y., Okano HJ, Imai T., Poole AJ., Suzuki N., Keirstead HS and Okano H.
  • Journal Title: Neurology and Clinical Neuroscience Volume: 1 Issue: 1 Pages: 24-31
  • DOI: 10.1002/ncn3.2
  • Peer Reviewed
• [Journal Article] RNA結合タンパク質による遺伝子制御と神経発生・神経機能 (2012)
  • Author(s): 今井貴雄、岡野栄之
  • Journal Title: 細胞工学6月号特集「神経細胞特異的な翻訳制御：鍵を握るRNA結合タンパク質/mRNA輸送/non-coding RNA」編者：武井延之 Volume: 31 Pages: 668-674
• [Presentation] Musashi1 Post-Transcriptionally Regulates The Gene Expression In Neural Stem/Progenitor Cells And Glioma. (2012)
  • Author(s): Imai T., Muto J., Kawase S., Shibata S., Okano H.
  • Organizer: The 4th Biennial Symposium on Brain and Mind Research in the Asia-Pacific (BMAP 2012)/The FIRST Program ‘Strategic Exploitation of Neuro-Genetics for Emergence of the Mind’ Symposium (FIRST 2012)
  • Place of Presentation: North Hall, Mita Campus, Keio University, Tokyo, Japan
• [Presentation] Musashi-1による胃特異的なm-Numb翻訳促進作用を介した胃粘膜修復機構
  • Author(s): 高橋哲史, 鈴木秀和, 今井貴雄, 土本寛二, 岡野栄之, 日比紀文
  • Organizer: 第40回日本潰瘍学会/第14回国際潰瘍学会
  • Place of Presentation: 東京、京王プラザホテル
• [Presentation] MUSASHI1 POST-TRANSCRIPTIONALLY REGULATES THE GENE EXPRESSION IN NEURAL STEM/PROGENITOR CELLS AND GLIOMA.
  • Author(s): Imai T., Muto J., Kawase S., Shibata S., Okano H.
  • Organizer: International Society for Stem Cell Research (ISSCR) 10th Annual Meeting
  • Place of Presentation: Pacifico Yokohama Conference Center, Yokohama, Japan
• [Presentation] Musashi1 post-transcriptionally regulates the gene expression in neural stem/progenitor cells and glioma
  • Author(s): 今井 貴雄, 武藤 淳, 河瀬 聡, 芝田 晋介, 岡野 栄之
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡、福岡国際会議場・マリンメッセ福岡