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1991 Fiscal Year Final Research Report Summary

Development of adeno-associated virus (AAV) vector

Research Project

Project/Area Number 02454181
Research Category

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

Allocation TypeSingle-year Grants
Research Field Virology
Research InstitutionTokyo Institute of Technology (1991)
The University of Tokyo (1990)

Principal Investigator

HANDA Hiroshi  Tokyo Institute of Technology, Faculty of Bioscience and Biotechnology, Professor, 生命理工学部, 教授 (80107432)

Project Period (FY) 1990 – 1991
KeywordsAdeno-associated virus / Virus vector / Envelope protein / Adenovirus / Helper virus / Regulation of expression / Transcription factor / Affinity-latex particles
Research Abstract

The human parvovirus adeno-associated virus (AAV) grows in the presence of helper functions provided by helper adenovirus. AAV is not pathogenic to human beings. When AAV infects cells in the absence of a helper virus, it frequently integrates its genome into the host cell genome. AAV has several advantages as virus vectors. To exploit AAV as virus vectors, we have worked on three main projects, (1) reconstruction of AAV genome, (2) reconstruction of helper adenovirus genome, (3) regulation of expression of foreign genes inserted into AAV genome.
(1) To reconstruct AAV genome, we prepared a recombinant plasmid pAAV which contained whole AAV genome. Several foreign gents, were inserted into the envelope protein region of PAAV. Human thymus epithelial cells were immortalized with the plasmid pAAV-SV40 DNA, which contained the SV40 early gene in the envelope region of PAAV. It indicated that the AAV vector was able to be developed from the recombinant plasmid.
(2) We also made a recombinant helper adenovirus, which expressed AAV envelope proteins to complement the defectiveness of recombinant AAV vectors. We have tried to produce recombinant AAV vectors by co-introduction of pAAV-SV40 DNA and the recombinant helper adenovirus into 293 cells. The recombinant AAV virus was obtained but the virus titer was low, because of low efficiency of production of AAV envelope proteins.
(3) To study the regulation of gene expression of foreign genes inserted into AAV genome, we have developed the affinity latex particles to purify DNA-binding transcription factors. By using the particles, we were able to purify multiple members of the ATF/E4TF3 family or two subunits of E4TF1 directly from crude cell extracts. The particles would be useful for studying various transcription factors.
On the basis of this study, we will try further to exploit AAV as useful virus vectors.

  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Tadashi Wada: "Different biological activities of the hetero- and homodimers formed by the 47kd and 43kd proteins of transcription factor ATF/E4TF3" J.Virol.65. 557-564 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mizuko Mizutani: "Negative supercoiling of DNA facilitates an interaction between transcription factor IID and the fibroin gene promoter" Proc.Natl.Acad.Sci.USA. 88. 718-722 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shin Aizawa: "Establishment of a variety of human bone marrow stromal cell lines by the recombinant SV40-adenovirus vector" J.Cell.Physiol.148. 245-251 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yoshihiro Usuda: "Affinity-purification of transcription factor IIA from HeLa cell nuclear extracts" EMBO J.10. 2305-2310 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sadaki Inokuchi: "Immortalization of human epidermal keratinocytes by the recombinant SV40-adenovirus vector" In Vitro Cell.Dev.Biol.27. 827-828 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshifumi Hibi: "Establishment of epithelial lines from human and mouse thymus immortalized by the 12S adenoviral E1A gene product" Thymus. 18. 155-167 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 青山 友三他: "ウイルス感染症の臨床と病理" 医学書院, 247 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tadashi Wada: "Different Biological activites of the hetero-and homodimers formed by the 47kd and 43kd proteins of transcription factor ATF/E4TF3" J. Virol.65. 557-564 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mizuko Mizutani: "Negative supercoiling of DNA facilitates an interaction between transcription factor IID and the fibroin gene promoter" Proc. Natl. Acad. Sci. USA. 88. 718-722 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shin Aizawa: "Establishment of a variety of human bone marrow stromal cell lines by the recombinant SV40-adenoviru vector" J. Cell. Physiol. 245-251 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshihiro Usuda: "Affinity-purification of transcription factor IIA from HeLa cell nuclear extracts" EMBO J.10. 2305-2310 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sadaki Inokuchi: "Immortalization of human epidermal keratinocytes by the recombinant SV40-adenovirus vector" In Vitro Cell. Dev. Biol.27. 827-828 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toshifumi Hibi: "Establishment of epithelial lines from human and mouse thymus immortalized by the 12S adenoviral ElA gene product" Thymus. 18. 155-167 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yuzo Aoyama: "Clinics and Pathology of Virus Infectious Diseases" Igakushoin. (1991)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-16  

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