| Project/Area Number |
08557065
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Hematology
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| Research Institution | Osaka University |
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Principal Investigator |
TAKEDA Junji Osaka University Medical School Professor, 医学部, 教授 (50163407)
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| Co-Investigator(Kenkyū-buntansha) |
SAKATA Tsuneaki DNAVEC Research inc.Chief Researcher, 室長
NAKANISHI Mahito Osaka University Research Institute for Microbial Deseases Assosiate Professor, 微生物病研究所, 助教授 (10172355)
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| Project Period (FY) |
1996 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥16,100,000 (Direct Cost: ¥16,100,000)
Fiscal Year 1998: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1997: ¥4,400,000 (Direct Cost: ¥4,400,000)
Fiscal Year 1996: ¥7,700,000 (Direct Cost: ¥7,700,000)
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| Keywords | Cre / loxp / Hematopoietic stem cell / Tissue specific gene targeting / Pig-a / Pig-a / コンディショナルジーンターデッティング / 膜融合リポソーム / コンディショナルジーンターゲティング |
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| Research Abstract |
Gene disruption in murine embryonic stem (ES) cells allows to study functions of endogenous genes after establishing mice derived from targeted ES cells. Since many gene disruptions cause embryonic lethality, it is not possible to analyze specific gene functions in adult mice. To circumvent these difficulties, we applied Cre/loxP system. The system allows to disrupt a gene flanked by identically oriented loxP sites with Cre-expression-dependent manner.
In this study, we tried to apply two methods to introduce Cre recombinase into mouse genome, exogenous and endogenous way. For the former, liposome containing Gre was produced. Addition of the liposome to hematopoietic cells in vitro resulted in disruption of loxP containing gene (Pig-a). However, transfer of these cells into lethally irradiated mice revealed that Pig-a disruption did not occur in the hematopoietic stem cells. This result suggest that the exogenous Gre by the liposome has limitation to introduce Cre into the genome. For the latter, we established a series of Gre transgenic mice. Cre transgene worked very efficiently including hematopoietic stem cells.
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