| Project/Area Number |
08457275
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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 |
Hematology
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| Research Institution | Niigata University |
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Principal Investigator |
TAKAHASHI Masuhiro Niigata University, College of Biomedical Technology, Department of Medical Technology, Professor, 医療技術短期大学部, 教授 (90179531)
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| Co-Investigator(Kenkyū-buntansha) |
FURUKAWA Tatsuo Niigata University, School of Medicine, Division of Bioclean Room, Associate Pro, 医学部・附属病院, 助教授 (00272849)
KOIKE Tadashi Niigata University, School of Medicine, Division of Blood Transfusion, Associate, 医学部・附属病院, 助教授 (30170161)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1997: ¥700,000 (Direct Cost: ¥700,000)
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| Keywords | homologous recombination / bcr-abl / thymidine kinase / neo / gene replacement / bvr-abl融合遺伝子 / homologous recombination / leukemia / gene therapy / BCR-ABL oncogane / RT-PCR / Souther Blotting / Neo / Thymidine kinase |
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| Research Abstract |
Ideal somatic gene therapy for genetic diseases is to treat by the replacement of an abnormal gene with a normal gene and to normalize the characteristics of abnormal cells. Up to now, repair of an abnormal gene can be performed only by homologous recombination between that abnormal gene and the gene for correction. Our study was designed to discover whether homologous recombination occurs in hematopoietic cells and if is possible to establish gene therapy using homologous recombination. Murine hematopoietic cell line, FDC-P2, which was factor-dependent but became factor-independent by being transfected with p210bcr-abl was used for the target cell for homologous recombination. The plasmid for homologous recombination was made by inserting neo-gene in the region of bcr-abl, thymidine kinase-gene in the region outside of bcr-abl, and placing bcr-abl region reversely for defining the homologous recombination region to bcr-abl. Transfection was done by electroporation and selection was performed by adding G418 and gancyclovir. One clone was established and confirmed to be replaced by the plasmid made for homologous recombination by RT-PCR findings of thymidine kinase (-), neo (+), bcr-abl (-) and a reasonable change of thr band by Southern blot analysis. The clone became factor-independent. These findings suggest the possibility of homologous recombination-associated gene replacement therapy for hematological diseases including leukemia.
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