| Project/Area Number |
14360174
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TOJO Hideaki The University of Tokyo, Graduate School of Agricultural and Life Sciences, Professor, 大学院・農学生命科学研究科, 教授 (20041668)
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| Co-Investigator(Kenkyū-buntansha) |
NAITO Kunihiko The University of Tokyo, Graduate School of Agricultural and Life Sciences, Associate Professor, 大学院・農学生命科学研究科, 助教授 (20188858)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥14,700,000 (Direct Cost: ¥14,700,000)
Fiscal Year 2003: ¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 2002: ¥8,200,000 (Direct Cost: ¥8,200,000)
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| Keywords | Cre / Pseudo loxP / IoxP / Transgenic mice / DNA micro injection / Gene transfer / トランスジェニック / マウス / loxp / Cre酵素 / Pseudoloxp / psedoloxp / CAG |
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| Research Abstract |
Functional lox-like sequences have been identified within the yeast and mammalian genome. These hetero-specific lox sites also allow Cre recombinase to specifically target efficient integration of exogenous DNA into the endogenous pseudo-IoxP (ΨLoxP) sequences that occur naturally in the host genorne using the Cre/loxP integrative recombination system. We investigated whether the Cre/ΨloxP system is useful for site-specific integration of a transgene and for improving the production efficiency of transgenic animals. This is the first report on Cre-mediated integrative recombination targeting an endocienaus lox-like sequence termed pseudo-loxm5 (ΨLoxm5 ) in early mouse embryos. First we characterized the Cre/Ψloxm5 system in the embryonic environment. Cre expressing plasmid and a transclene (CMV/LacZ gene) flanked by two pseudo loxm5 sites were co-microinjected into the pronucleus of fertilized mouse oocytes. The injected eggs were transferred into foster mothers and recombination products were investigated. The results show that the Ψbloxm5 site is an active substrate far Cre mediated recombination in the mouse embryonic environment. The transgenesis efficiency was up to 27% (6/22). The site specific integration of the transgene into the endogenous ΨLoxm5 site was found in the transgenic pups. Our findings demonstrated thai the Cre/Ψloxm5 integrative recombination system was an efficient and simple strategy for targeting an endogenous lox-like site in mammalian embryos.
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