1997 Fiscal Year Final Research Report Summary
Basic research regarding gene therapy for leukemia by using homologous recombination
| 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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| Keywords | homologous recombination / bcr-abl / thymidine kinase / neo / gene replacement |
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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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Research Products
(13 results)
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[Publications] Takahashi M,Shigeno N,Takahashi H,Suzuki N,Masuko M,Nikkuni K,Toba K,Furukawa T,Aoki S,Kishi K,Koike T,Aizawa Y: "Effects of transfection of P210bcr-abl and bcr-v-abl into the factor-dependent human leukemia cell line HSM-911" Leuk Res. 21 (11/12). 1115-1123 (1997)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Toba K,Koike T,Takahashi M,Kishi K,Hashimoto S,Takahashi H,Uesugi Y,Ishikawa T,Aoki S,Maeo S,Naito M,Aizawa Y,Shibata A: "Characterization and sensitivity to interleukin-2 and interferon-alpha of leukemic cells from a patient with large granular lymphocytic leukemia associated with chronic active Epstein-Barr virus infection" Leuk Res. 21 (10). 941-950 (1997)
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[Publications] Takahashi M,Shigeno N,Suzuki N,Takahashi H,Nikkuni K,Masuko M,Sakaue M,Furukawa T,Kishi K,Koike T,Aizawa Y: "Confiscation of Autonomous Proliferation from P210bcr-abl Transfected Cells by Re-transfection with DNA for Homologous Recombination" Transgenics. (in press).
Description
「研究成果報告書概要(欧文)」より
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