Generation of synthetic rescue system in murine haploid ES cells.
| Project/Area Number |
26640124
|
|---|
| Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Medical genome science
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
TAKEDA Junji 大阪大学, 医学(系)研究科(研究院), 教授 (50163407)
|
|---|
| Project Period (FY) |
2014-04-01 – 2015-03-31
|
| Project Status |
Completed (Fiscal Year 2014)
|
| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2014: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
|
|---|
| Keywords | ゲノムワイド関連解析 / ハプロイドES細胞 |
|---|
| Outline of Final Research Achievements |
We performed experiments to generate a novel synthetic rescue system in murine haploid ES cells. For conditional KO system in haploid ES cells, we first tried to insert tamoxifen- inducible cassette, ERT2-iCre-ERT2 into the Rosa 26 locus. Since homologous targeting in haploid ES cells was very low, we used ZFN for the targeting.
Meanwhile, production of CRISPR-gRNA library has bee reported. The library is able to disrupt both alleles of a gene of interest.
We modified both alleles of Oct4 to floxed ones and we introduced CRISPR-gRNA library to disrupt endogenous genes randomly. After disruption of Oct4 by activation of Cre with tamoxifen. We isolated undifferentiated clones. We are analyzing these clones right now.
|
Report
(2 results)
Research Products
(4 results)
