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Basic study on development of transgenic farm animals for medical use by using RNA interference

Research Project

Project/Area Number 16380197
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Basic veterinary science/Basic zootechnical science
Research InstitutionThe University of Tokyo

Principal Investigator

TOJO Hideaki  The University of Tokyo, Graduate School of Agricultural and Life Sciences, Professor, 大学院農学生命科学研究科, 教授 (20041668)

Co-Investigator(Kenkyū-buntansha) NAITO Kunihiko  The University of Tokyo, Graduate School of Agricultural and Life Sciences, Associate Professor, 大学院農学生命科学研究科, 助教授 (20188858)
NARUSE Katsutoshi  The University of Tokyo, Graduate School of Medicine, Assistant, 大学院医学研究科, 助手 (50291323)
KIKUCHI Kazuhiro  National Institute of Agrobiological Sciences, Chief Investigator, 主任研究官 (20360456)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥15,600,000 (Direct Cost: ¥15,600,000)
Fiscal Year 2005: ¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 2004: ¥8,800,000 (Direct Cost: ¥8,800,000)
KeywordsGene knock down / Gene manipulated farm animals / Medical use-transgenic farm animals / Galactose transferase / RNA interference / RNAi / トランスジェニック / ブタ卵子 / α-1,3-Gal / EGFP / トランスジェニックブタ
Research Abstract

1)For production of transgenic pigs, there are many problems such as preparation of fertilized eggs, in vitro development of embryos and so on. As basic study on elevating the production efficiency of fertilized eggs, we have investigated molecular mechanism of development of oocytes during maturation. To examine whether the GV is required for MPF activation in mammalian oocytes, porcine immature oocytes were enucleated and their MPF activity and CCNB levels were investigated. The activation of MPF at the start of maturation was detected at normal levels in enucleated oocytes, whereas reactivation to induce the second meiosis was not observed. Although protein synthesis was found to be normal both qualitatively and quantitatively, even in the absence of the nucleus, CCNB1 did not sufficiently accumulate in the enucleated oocytes. The defects in the enucleated oocytes were reversed by the injection of GV material into the enucleated oocytes. Furthermore, the inhibition of CCNB1 degradation revealed drastic accumulation of CCNB1, indicating active synthesis of CCNB1 in enucleated oocytes. The mitogen activated protein kinase cascade remained unaffected by enucleation. These results indicate that GV is not required for the activation of MPF during the first meiosis, but that it is required for the second meiosis because of its promotion of CCNB1 accumulation. 2)In this study, SLPI(secretory leukocyte protease inhibitor) that has been well documented and is familiar protein in our laboratory was used for targeting RNA interference(RNAi). We have studied the effect of RNAi of SLPI on the proliferation of human cancer cells, MCF-7 and MDA-MB-231. We have not yet obtained satisfactory results of si RNA construct effective for RNAi. We are attempting to design more effective structure of siRNA of SLPI for RNAi

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (23 results)

All 2006 2005 2004

All Journal Article (20 results) Book (3 results)

  • [Journal Article] Multifunctional roles of MT1-MMP in myofiber formation and morphostatic maintenance of skeletal muscle2006

    • Author(s)
      Ohtake Y
    • Journal Title

      J Cell Sci. 119

      Pages: 3822-3832

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Analysis of the promoter activity of the mutated whey acidic protein(WAP) gene2006

    • Author(s)
      Nukumi Y
    • Journal Title

      J Reprod Dev. 52

      Pages: 315-320

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Investigation of the fate of Sry-expressing cells using an in vivo Cre/loxP system2006

    • Author(s)
      Ito M
    • Journal Title

      Dev Growth Differ 48

      Pages: 41-47

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Activities of maturation-promoting factor(MPF) and mitogen-activated protein kinase(MAPK) are not required for the global histone deacetylation.2006

    • Author(s)
      Endo T
    • Journal Title

      Reproduction 131

      Pages: 439-447

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Multifunctional roles of MT1-MMP in myofiber formation and morphostatic maintenance of skeletal muscle2006

    • Author(s)
      Ohtake Y.
    • Journal Title

      J Cell Sci. 119

      Pages: 3822-3832

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Analysis of the promoter activity of the mutated whey acidic protein (WAP) gene2006

    • Author(s)
      Nukumi N.
    • Journal Title

      Reprod Dev. 52

      Pages: 315-320

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Investigation of the fate of Sry-expressing cells using an in vivo Cre/loxP system2006

    • Author(s)
      Itoh M
    • Journal Title

      Dev Growth Differ

      Pages: 41-4748

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Activities of maturation-promoting factor (MPF) and mitogen-activated protein kinase (MAPK) are not required for the global histone deacetylation2006

    • Author(s)
      Endo T.
    • Journal Title

      Reproduction 131

      Pages: 439-447

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Study of germinal vesicle requirement for the normal kinetics of maturation/M-phase-promoting factor activity during porcine oocyte2006

    • Author(s)
      Sugiura K.
    • Journal Title

      Biol Reprod 74

      Pages: 593-600

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Study of germinal vesicle requirement for the normal kinetics of maturation/M-phase-promoting factor activity during porcine oocyte2005

    • Author(s)
      Sugiura K
    • Journal Title

      Biol Reprod 174

      Pages: 593-600

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Evaluation of H-FABP as a marker of ongoing myocardial damage using hGH transgenic mice2005

    • Author(s)
      Naraoka H
    • Journal Title

      Clinica Chimica Acta 361

      Pages: 123-128

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Evaluation of H-FABP as a marker of ongoing myocardial damage using hGH transgenic mice2005

    • Author(s)
      Naraolka H.
    • Journal Title

      Clinica Chimica Acta 361

      Pages: 123-128

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Aberrant development of mammary glands but precocious expression of beta-casein in transgenic females ubiquitously expressing WAP transgene2005

    • Author(s)
      Iwamori K
    • Journal Title

      J Reprod Dev 51

      Pages: 579-592

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Cdk2 activity is essential for the first to second meiosis transition in porcine oocytes2005

    • Author(s)
      Sugiura K
    • Journal Title

      J Reprod Dev 51

      Pages: 143-149

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Chanegs in histone modifications during in vitro maturation of porcine oocytes2005

    • Author(s)
      Endo T
    • Journal Title

      Mol Reprod Dev 71

      Pages: 123-128

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Detection of elements responsible for stage- and tissue-specific expression of mouse Sry using an in vitro Cre/loxP system2005

    • Author(s)
      Ito M
    • Journal Title

      Biochem Biophys Res Commun 337

      Pages: 264-270

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Expression and localization of matrix metalloproteinases (MT1-MMP, MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) during synepitheliochorial placentation of goats (Capra hircus)2004

    • Author(s)
      Uekita T
    • Journal Title

      Placenta 25

      Pages: 810-819

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Inhibitory function of whey acidic protein in the cell-cycle progression of mouse mammary epithelial cells (EpH4/K6 cells)2004

    • Author(s)
      Ikeda K
    • Journal Title

      Journal of Reproduction and Development 50

      Pages: 87-96

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Analysis of the roles of cyclin B1 and cyclin B2 in porcine oocyte maturation by inhibiting synthesis with antisense RNA injection.2004

    • Author(s)
      Kuroda T
    • Journal Title

      Biology of Reproduction 70

      Pages: 154-159

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Regulatory function of whey acidic protein in the proliferation of mouse mammary epithelial cells in vivo and in vitro.2004

    • Author(s)
      Nukumi N
    • Journal Title

      Developmental Biology 274

      Pages: 31-44

    • Related Report
      2004 Annual Research Report
  • [Book] 図説 基礎動物生理学2006

    • Author(s)
      東條 英昭
    • Total Pages
      160
    • Publisher
      (株)アドスリー
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] Illustrated Basic Animal Physiology2006

    • Author(s)
      Hideaki Tojo
    • Total Pages
      160
    • Publisher
      Adthree, Tokyo
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] シリーズ応用動物科学/バイオサイエンス「トランスジェニック動物」2004

    • Author(s)
      東條 英昭
    • Total Pages
      141
    • Publisher
      朝倉書店
    • Related Report
      2004 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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