| 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 *help |
¥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.
|
