1994 Fiscal Year Final Research Report Summary
Production of mouse models for human diseases by gene targeting
| Project/Area Number |
04044136
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Institute of Molecular Embryology and Genetics Kumamoto University School of Medicine |
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Principal Investigator |
YAMAMURA Ken-ichi Kumamoto University School of Medicine, 医学部, 教授 (90115197)
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| Co-Investigator(Kenkyū-buntansha) |
SMITHIES Oliver North Carolina State University, School of Medicine, 医学部, 教授
SOLTER Davor Max-Planck Institute for Immunobiology, 発生部門, 部門長
KANAME Tadashi Kumamoto University School of Medicine, 医学部, 助手 (40264288)
ARAKI Kimi Kumamoto University School of Medicine, 医学部, 助手 (90211705)
ABE Kuniya Kumamoto University School of Medicine, 医学部, 助教授 (40240915)
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| Project Period (FY) |
1992 – 1994
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| Keywords | Gene targeting / homologous recombination / gene trap / ES cell / transgenic mouse / yeast artificial chromosome |
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| Research Abstract |
The purpose of this project is to discuss about the possibility and application of gene targeting techniques in biomedical research. We visited Dr.Smithies, North Carolina State University, to discuss about the recent advances in transgenic technology. Chimera production by aggregating ES cells with morula was shown to be used for only limited cell line, such as R1 developed by Nagy. It is now possible to produce transgenic mice by introducing YAC DNA containing up to 1 megabase DNA.The size of YAC DNA treated with solution containing polyamine was demonstrated to be smaller than the inner diameter of injection pipette used for microinjection of DNA into pronuclei of fertilized eggs. Thus, the microinjection method was most feasible approach to produce YAC transgenic mice. As the contamination of agarose results in the breakdown of giant YAC DNA,it is essential to dissolve agarose completely by agarase before isolation of DNA.
We visited Dr.Solter, Max-Planck-Institute for lmmunobiology, to discuss about the gene trap method. It is important to develop a new screening system to identify the genes involved in the formation of germ layr. Thus, we cultured ES cells in LIF-free medium. These ES Cells aggregated within a few days and developed into embryoid body. Liver-specific genes, such as transthyretin gene or, albumin gene, were expressed in a similar pattern observed during liver cell differentiation. Bacteriophage recombination system, Cre-loxP,is shown to be useful to knock out the gene in a tissue-specific and developmentally specific manner. It was also suggested that the recombinase, Cre, should be expressed in a nucleus to exert its effect efficiently.
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Research Products
(12 results)
