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Production of transgenic livestock animals bearing antisense growth hormone cDNA and analysis of mechanisms of the transgene expression

Research Project

Project/Area Number 05304022
Research Category

Grant-in-Aid for Co-operative Research (A)

Allocation TypeSingle-year Grants
Research Field Applied animal science
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

YAMADA Masayasu  Kyoto Univ.College of Agri., Associate Professor, 農学部, 助教授 (10243073)

Co-Investigator(Kenkyū-buntansha) SAKAI Hiroshi  Kyoto Univ.College of Agri.Associate professor, 農学部, 助教授 (60089117)
IRITANI Akira  Kinki Univ.Res.Inst.of Bio.-Oriented Sci.and Tech.Professor, 生物理工学部, 教授 (80026385)
YAMANO Yosiaki  Tottori Univ.Faculty of Agri., Associate Professor, 農学部, 助教授 (00182593)
WAKASUGI Noboru  Nagoya Univ.School of Agri., Professor, 農学部, 教授 (00023490)
YAMAMURA Kenichi  Kumamoto Univ.School of Med., Professor, 医学部, 教授 (90115197)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1994: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥3,500,000 (Direct Cost: ¥3,500,000)
KeywordsGene transfer / Miniature livestock animals / Antisense growth hormone gene / Tissue-specific expression / In vitro fertilization of goat / ニワトリIGF-1遺伝子 / 遺伝子発現機構 / 矮性化遺伝子 / IGF-1レセプター遺伝子 / ジーンターゲッティング
Research Abstract

In an attempt to produce transgenic rats bearing antisense DNA complementary to sequences in the rat growth hormone(GH)gege and to investigate whether antisense RNA expression can modulate GH gene expression and body growth , we produced four transgenic founder rats(F_0)by introducing into rat embryos a fusion gene, which consisted of rat GH promoter containing with four copies of thyroid hormone response element(TRE)and antisense cDNA sequences for rat GH.This transgene promoter directed 2.8-fold stimulation of CAT gene expression in transfected GH_3 rat pituitary tumor cells compared with the rat GH promoter alone.Two of four transgenic rats expressed strongly antisense RNA in the pituitary and lymphoid tissues.Transgenicoffspring(F_1)from each founder rat exhibited dwarfism at as early as 3-4 weeks of age, and they exhibited 70-80% reduced growth rate compared with their nontransgenic littermates.
We next examined the effects of inserting tandem copies of TRE on tissue-specific expression of antisense GH RNA and modulation of body growth in the transgenic rats.When antisense cDNA for rat GH linked to rat GH promoter alone was introduced into rat embryos, the offspring(F1)of the transgenic fonder rats did not exhibited dwarfism, whereas expression of antisense RNA was detected in their various tissues including pituitary and lymphoid tissues using RT-PCR.From the results, it is suggested that TRE is necessary for the suppression of GH gene expression by antisense RNA.We are now trying to produce a miniature goat by inhibiting GH gene expresion with introduction of antisense goat GH cDNA into pronuclear embryos derived from IVM/IVF oocytes.
We have also demonstrated that a 6-kb upstream region of the human transthyretin gene retained all the cis-elements required for developmental, tissue-specific, and quantitatively normal expression.

Report

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

    (28 results)

All Other

All Publications (28 results)

  • [Publications] K.Matsumoto: "Growth retardation in rats whose growth hormone gene expression was suppressed by antisense RNA transgene." Mol.Reprod.Dev.36. 53-58 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Nagata: "A 6-Kb upstream region of the numan transthyretin gene can direct developmental,tissue-specific,and quantitatively normal expression in transgenic mouse" J.Biochem. 117. 169-175 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.S.Kim.: "Effects of bovine follicular fluld on invitro maturation of bovine oocytes" Jpn.J.Embryo Transfer. 17. 1-5 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Yamamura: "Transgenic mouse model for human genetic diseases" Mol.Reprod.Dev.36. 248-250 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] S.Mikawa: "Dynamic aspects in the expression of the goat insulin-like growth factor-I(IGF-I)gene:diversity in transcription and post-transcription." Biosci.Biotech.Biochem.59. 87-92 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Tsudzuki: "Allelisin of panda and dotted white plumage genes in Japanese quail" J.Heredity. 83. 226-229 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Tsudzuki: "Dotted white-A plumage color mutant in Japanese quail(Coturnix japonica)" Exp.Anim.41. 25-31 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] A.Fujimoto: "A tail length modifier gene discovered in Japanese wild mice(Mus musculus molossinus)." Jpn.J.Genet.66. 141-154 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Yamano: "Cloning of cDNAs for cecropins A and B,and expression of the genes in the silkworm,Bombyx mori." Biosci.Biotech.Biochem.58. 1476-1478 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] I.Morishima: "Lysozyme activity in immunized and non-immunized hemolymph during the development of the silkworm,Bombyx mori." Comp.Biochem.Physiol.108. 311-314 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mastumoto K.: "Growth retardation in rats whose growth hormone gene expression was suppressed by antisense RNA transgene." Mol.Reprod.Dev.36. 53-58 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nagata, Y.: "A 6-kb upstream region of human transthyretin gene can direct developmental, tissue-specific, and quantitatively normal expression in transgenic mouse." J.Biochem.117. 169-175 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yamamura, K.: "Trasgenic mouse model for humangenetic diseases." Mol.Reprod.Dev.36. 248-250 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Tsudzuki, M.: "Allelism of panda and dotted white plumage genes in Japanese quail." J.Heredity. 83. 226-229 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Tsudzuki, M.: "Dotted white-A plumage color mutant in Japanese quail(Coturnix japonica)." Exp.Anim.41. 25-31 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujimoto, A.: "A tail length modifier gene discovered in the Japanese wild mice(Mus musculus molossinus)." Jpn.J.Genet. 66. 141-154 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mikawa, S.: "Dynamic aspects in the expression of the goat insulin-like growth factor-1(IGF-1)gene : diversity in transcription and post-transcription." Biosci.Biotech.Biochem.59. 87-92 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yamano, Y.: "Cloning of cDNAs for cecropins A and B,and expression of the genes in the silkworm, Bombyx mori." Biosci.Biotech.Biochem.58. 1476-1478 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Morisima, I.: "Lysozyme activity in immunized and non-immunized hemolymph during the development of the silkworm, Bombyx mori." Comp.Biochem.Physiol.108. 311-314 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kim, K.S.: "Effects of bovine follicular fluid on in vitro maturation of bovine oocytes." Jpn.J.Embryo Transfer. 17. 1-5 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yamada, M.: "Role of nuclear pore complex in simian virus 40 nuclear targeting." J.Virol.67. 119-130 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.S.KIM: "Effects of bovine follicular fluid on in vitro maturation of bovine oocytes" Jpn.J.Embryo Transfer. 17. 1-5 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] N.ISHII: "Functional complementation of nuclear targeting-defective mutants of Simian virus 40 structural proteins" J.Virol.68. 8209-8216 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Y.NAGATA: "A 6-kb upstream region of the human transthyretin gene can direct developmental,tissue-specific,and quantatively normal experssion in transgenic mouse" J.Biochem.117. 169-175 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] S.MIKAWA: "Dynamic aspects in the expression of the goat insulin-like growth factor-1(IGF-1)gene:Diversity in transcription and post-transcription" Biosci.Biotech.Biochem.59. 87-92 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.MATSUMOTO: "Growth Retardation in Rats Whose Growth Hormone Gene Expression Was Suppressed by Antisense RNA Transgene" Molecular Reproduction and Development. 36. 53-58 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.TSUZUKI: "Allelism of Panda and Dotted White Plumage Genes in Japanese Quail" Journal of Heredity. 84. 226-229 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] K,-I,YAMAMURA: "Transgenic Mouse Model for Human Genetic Diseases" Molecular Reproduction and Development. 36. 248-250 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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