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

2014 Fiscal Year Final Research Report

• 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
• 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

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.

Free Research Field

分子生物学、幹細胞生物学