2000 Fiscal Year Final Research Report Summary
Establishment of genetically modified mouse libraries using Cre/loxP gene trap
| Project/Area Number |
11558098
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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 | 展開研究 |
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| Research Field |
Laboratory animal science
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| Research Institution | Osaka University |
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Principal Investigator |
OKABE Masaru Osaka University, Genome Information Research Center, Professor, 遺伝情報実験施設, 教授 (30089875)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Hiroshi Chugai Pharmaceutical Co., Ltd., Exploratory Res.Lab., Chief researcher, 創薬資源研究所, 主任研究員
IKAWA Masahito Genome Information Research Center, Assistant Professor, 遺伝情報実験施設, 助手 (20304066)
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| Project Period (FY) |
1999 – 2000
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| Keywords | Cre recombinase / polyA trap / ES cells / Cre / IoxP system / GFP / green mouse |
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| Research Abstract |
The purpose of this study is to estimate the possibility of establishing genetically modified mouse libraries using Cre/lop trap system. The research also aimed to produce transgenic mouse lines that express Cre recombinase in various ways to enable a second generation of gene knockout.
1) Gene trap in ES cells and identification of the trapped gene
We have produced trap vector that contains polyA less Puromycin resistant gene under the PGK promoter. At the same time, the vector was designed to trap a promoter of a certain endogenous gene and express Cre recombinase. Using this vector, we have obtained 600 trapped clones. After the analysis of the trapped clones using 3'-RACE, we identified 9 known gene 2 EST sequences. Other sequences were originated from unknown genes. The vector was shown to be effective to trap genes which are silent in ES cells.
2) Prodction of Chimeric mouse from trapped ES cells.
Chimeric mice are produced from the trapped ES cell lines and at the present more than 10 genetically modified mouse lines are established. The phenotypes of these gene-disrupted mice are now under investigations. The expression of the Cre recombinase from these trapped mice are also examined using GFP transgenic mice which becomes green in organs where Cre is expressed.
In order to identify the localization of Cre expressing organs (or cells), we produced reporter "green mice" which turns fluorescent green when Cre recombinase recombine the transgene.
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