Attempt to establish totipotent stem cells using degron system
| Project/Area Number |
16K14594
|
|---|
| Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Laboratory animal science
|
|---|
| Research Institution | Kyoto University |
|---|
Principal Investigator |
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
成瀬 智恵 京都大学, 医学研究科, 准教授 (30372486)
浅野 雅秀 京都大学, 医学研究科, 教授 (50251450)
|
|---|
| Project Period (FY) |
2016-04-01 – 2019-03-31
|
| Project Status |
Completed (Fiscal Year 2018)
|
| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
|
|---|
| Keywords | デグロン / マウス / ゲノム編集 / マウス初期胚発生 / デグロンシステム / 発生 / 初期胚 / 発生・分化 / 動物 |
|---|
| Outline of Final Research Achievements |
From the gene expression database, we selected candidate genes that were strongly expressed from the 8-cell stage to the morula stage, and confirmed 5 genes using early embryos and quantitative RT-PCR. In order to develop a gene transfer system for expressing these genes from one cell stage, we tried electroporation. So far, only one report has been reported that double-stranded DNA could be introduced into fertilized eggs by electroporation, and there has been no case to follow up. Condition examination was conducted independently, and the condition where expression of the marker gene could be confirmed stably in about 10-20% embryos were determined. In addition, we found a condition that allows 8-cell stage embryos to grow infinitely by suspension culture. Using these, it is expected that the condition closer to the early embryo can be created than the conventional method.
|
| Academic Significance and Societal Importance of the Research Achievements |
遺伝子翻訳産物を薬剤などにより自由に発現消去できる方法は現在限られており、新しい方法の開発が望まれている。目的のタンパク質にデグロンタグを付加することで、特定の薬剤により目的タンパク質を転写制御よりも素早く分解できる方法が実用化されれば、タンパク質の機能解析に大きく資することができる。また初期胚の8細胞期はES細胞よりもより幹細胞に近い全能性を持つと考えられており、8細胞期胚の性状を持ったまま増殖させられる条件を見つけることができれば、全能性の解明に資することができる。我々は候補遺伝子の特定と導入法の開発までたどり着くことができたが、この成果を用いて全能性幹細胞を確立したいと考えている。
|
Report
(4 results)
Research Products
(1 results)
