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THE STUDY ON RETURNING FROM LEUKEMIA BY USING HOMOLOGOUS RECOMBINATION

Research Project

Project/Area Number 05404040
Research Category

Grant-in-Aid for General Scientific Research (A)

Allocation TypeSingle-year Grants
Research Field Hematology
Research InstitutionNIIGATA UNIVERSITY

Principal Investigator

SHIBATA Akira  NIIGATA UNIVERSITY,SCHOOL OF MEDICINE,PROFESSOR, 医学部, 教授 (10004772)

Co-Investigator(Kenkyū-buntansha) KISHI Kenji  NIIGATA UNIVERSITY,SCHOOL OF MEDICINE,FELLOW, 医学部付属病院, 助手 (30186209)
KOIKE Tadashi  NIIGATA UNIVERSITY,SCHOOL OF MEDICINE,FELLOW, 医学部付属病院, 助手 (30170161)
TAKAHASHI Masuhiro  NIIGATA UNIVERSITY,SCHOOL OF MEDICINE,FELLOW, 医学部, 助手 (90179531)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥11,000,000 (Direct Cost: ¥11,000,000)
Fiscal Year 1994: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥8,900,000 (Direct Cost: ¥8,900,000)
KeywordsHOMOLOGOUS RECOMBINATION / GENE THERAPY / PGD210 / THYMIDINE KINASE / GANCICLOVIR / FDC-P2 / 脱白血病化 / BCR / ABL融合遺伝子 / 相同遺伝子組み替え / 電気パルス法 / Neo遺伝子 / HSV-tk遺伝子
Research Abstract

The essential gene therapy for malignant neoplasias including leukemia is to normalize the tumor cells by replacing the abnormal gene with normal relevant gene. In order to repair the abnormal oncogene, the procedure of homologous recombination is inevitable. We generated the plasmid for homologous recombination and performed the preliminary study using the plasmid. The plamid for homologous recombination (pGDTKbcrNeoablreverse) was generated by reversing the direction of bcr-Neo-abl (homologous region), which was obtained from pGD210 by inserting Neo gene, in the plasmid containing TK region, in order to make the homologous region more specific. Autonomous proliferation was acquired in the IL-3 dependent mouse hematopoictic cell line FDC-P2, which was transfected with bcr-abl gene. The copy number of transfected bcr-abl was one in all clones established. G418 resistance and the susceptibility for ganciclovir were observed in the FDC-P2 cells, which were transfected with pGDTKbcrNeoablreverse and demonstrated to express the messenger of Neo and TK.The present study suggested the applicability of the plasmid for knocking out the bcr-abl oncogene in the fashion of homologous recombination, since the cells with homologous recombination were demonstrated to be successfully selected by using G418 and ganciclovir. The pivotal issue of the gene introduction in this fashion is how to increase the rate of homologous recombination in the mammalian cells. For this purpose, the new strategy such as using Rad51 will be necessary to be developed.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report

Research Products

(13 results)

All Other

All Publications (13 results)

  • [Publications] Furukawa T.et al: "Establishment of a new cell line with the charactenistics" Leukemia. 8. 171-180 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Koike T.et al: "The therapeutic significance of mycloperoxidase" Leukemia. 8. 1253-1255 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Takahashi M.et al: "Transfection of the bcrlabloncogene into factor" Hematol Oncol. 12. 53-60 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Furukawa T,Koike T,Ying W,Kishi K,Aoki S,Gotoh T,Hashimoto S,Saitoh H,Hanano M,Shinada S,Moriyama Y,Takahashi M,Endo Y,Okano H,Maekawa T,Abe T,Shibata A: "Establishment of a new cell line with the characteristics of a multipotential progenitor from a patient with chronic myelogenous leukemia in early crythroblastic crisis." Leukemia. 8. 171-180 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Koike T,Takahashi M,Shibata A: "The therapcutic significance of myeloperoxidase detectable only by electron microscopy in acute leukemia." Leukemia. 8. 1253-1255 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Takahashi M,Furukawa T,Tanaka I,Ohsawa Y,Nikkuni K,Aoki A,Goto T,Hashimoto S,Kishi K,Koike T,Moriyama Y,Shibata A: "Transfection of the bcr/abl oncogene into factor-dependent cells by electroporation : Acquisitin of autonomous proliferation." Hematol Oncol. 12. 53-60 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Furukawa T.et al: "Establishment of a new cell line with the characteristics" Leukemia. 8. 171-180 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Koike T.et al: "The therapeutic significance of mycloperoxidase" Leukemia. 8. 1253-1255 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Takahashi M.et al: "Transfection of the bcrlabloncogene into factor" Hematol Oncol. 12. 53-60 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Takahashi M,et al: "Transfection of the bcr/abl oncogene into factor-dependent cells by electroporation:Acquisition of autonomous proliferation." Hematol Oncol.(in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Takahashi M,et al: "Gene introduction into granulocyte-macrophage progenitor cells by electroporation:The relationship between introduction efficiency and the proportion of cells in S-phase." Leuk Res.16. 751-767 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Takahashi M,et al: "Efficient introduction of a gene into hematopoietic cells in S-phase by electroporation." Exp Hematol.19. 343-346 (1991)

    • Related Report
      1993 Annual Research Report
  • [Publications] Takahashi M,et al: "Gene transfer into human leukemia cell lines by electroporation:experience with exponentially decaying and square wave pulse." Leuk Res.15. 507-513 (1991)

    • Related Report
      1993 Annual Research Report

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Published: 1993-03-31   Modified: 2016-04-21  

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