| Project/Area Number |
03670500
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pediatrics
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| Research Institution | Kumamoto University School of Medicine |
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Principal Investigator |
OHTANI Yoshinobu Kumamoto University School of Medicine, Department of Child Development, Assistant Professor, 医学部, 助教授 (10168982)
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| Co-Investigator(Kenkyū-buntansha) |
MIIKE Teruhisa Kumamoto University School of Medicine, Department of Child Development, Profess, 医学部, 教授 (90040617)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥700,000 (Direct Cost: ¥700,000)
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| Keywords | Duchenne muscular dystrophy / Animal model / Gene targeting / Embryonic stem cell / Dystrophin gene / Chimaera mouse / Enbryonic stem cell / Duchenne型筋ジストロフィ-症 |
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| Research Abstract |
Duchenne muscular dystrophy(DMD) is a X-linked recessive myopathy caused by mutation in the dystrophin gene(Dmd). The Dmd is transcribed in brain from a specific promoter that is different from the one used in muscle. The role of dystrophin in the muscle and brain is not clear. The mdx mouse mutant is due to mutation in the cognate murine Dmd. An animal model for DMD would be important for investigating its phenotypic diversity and pathogenesis and for evaluating therapeutic approaches.
We try to generate an animal model for DMD by creating a null allele in embryonic stem cells through gene targeting and using these genetically modified cells to establish a mouse strain carrying the mutation. The mutation is designed to generate null allele for Dmd lacking exon 1 of muscle type or brain type of it. The exon 1 of brain type was replaced with the E.coli lacZ gene by means of homologous recombintion in ES cells. The inclusion of the lacZ reporter may facilitate the monitoring of the temporal and spatial distribution through development. Utilizing a strategy of positive-negative selection together with analysis of individual clones by PCR-southern hybridization, we report here targeting frequency of brain type dystrophin locus with this construct of approximately 4/120 G418 resistant clones. In contrast, muscle type targeting clone have not been obtained yet. Chimaera carrying the mutation of brain type dystrophin locus is now being established.
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