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The establishment of prion protein humanaized mice using homologous recombination

Research Project

Project/Area Number 06404032
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Neurology
Research InstitutionTohoku University (1996)
Kyushu University (1994-1995)

Principal Investigator

KITAMOTO Tetsuyuki (1996)  Schoo, of Medicine, Tohoku University, professor, 医学部, 教授 (20192560)

立石 潤 (1994-1995)  九州大学, 医学部, 教授 (70033305)

Co-Investigator(Kenkyū-buntansha) NAKAMURA Kenji  Kyusyu University, assistant professor, 生体防御研究所, 助手 (90253533)
北本 哲之  東北大学, 医学部, 教授 (20192560)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥35,500,000 (Direct Cost: ¥35,500,000)
Fiscal Year 1996: ¥5,900,000 (Direct Cost: ¥5,900,000)
Fiscal Year 1995: ¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1994: ¥22,600,000 (Direct Cost: ¥22,600,000)
Keywordsprion protein / homologous recombination / site-specific recombination / humanized mice / transgenic mice / ヒト・プリオン蛋白 / 標的組換え / 部位特異的遺伝子組換え / クロイツフェルト・ヤコブ病 / プリオン蛋白 / 遺伝子置換
Research Abstract

To study human prions, we previously used the mice with wild type murine PrP gene for the transmission experiment. Mice have been used to do the transmission experiments from human or other animals with prion diseases. However, it takes long time for the successful transmission with the wild type mice, and the successful transmission was rarely observed in the wild type mice. Therefore, it is essential to establish humanized prion protein mice for the transmission experiment from human prion diseases. For this purpose, we designed the gene replacement method with the homologous recombination. Our method was based on the site-directed recombination using Cre-loxp system. At first, the loxp-neo-gpt-loxp cassette was used for the homologous recombination. One clone out of 140 G418-resistant clones was selected by the Southern blotting. We made the chimeta mice from this clone, and found out the germinal transmission from the chimera mice. The Cre plasmid was micro-injected in the fertiliz … More ed eggs from the heterozygous mice.
In 6 out 7 mice, the successful site-specific recombination was observed. We established the humanized mice using the homologous recombination and the site-specific recombination. In addition, we used the loxp-neo-loxp cassette for making other types of humanized mice. With this cassette, the efficient homologous recombination was obtained. One out of 60 clones or 3 out of 120 clones were recognized positively by the Southern blot analysis. At present, we are making the chimera mice from these ES cells.
For establishment of mice overexpressing human prion protein, we also made the transgenic mice. The natural promoter and the natural gene structure was obtained from the genomic library of the 129sv or I/ln mice. The transgene was constructed with the natural murine prion protein gene except for the exon 3. The exon 3 was replaced with human prion protein. Two constructs was micro-injected in the fertilized eggs from C57bl mice. We obtained 5 or 4 transgenic founder mice from each construct. These mice or germ-lined F1 mice showed a high expression of human prion protein mRNA.These replacement or transgenic mice were useful to analyze the human prion titer. Less

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (51 results)

All Other

All Publications (51 results)

  • [Publications] Kitamoto.T: "Human prion diseases with variant prion protein." Phil Trans R Soc Lond B. 343. 391〜398 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Inoue.I: "Japanese family with Creutzfeldt-Jakob disease with codon 200 point mutation of the prion protein gene." Neurology. 44. 299〜301 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yamada.S: "Creutzfeldt-Jakob disease transmitted by a cadaveric dura mater graft." Neurosurgery. 34. 704〜744 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Doi-Yi.R: "Distribution of prion protein in German patients with Creutzfeldt-Jakob disease is different from that in Japanese patients." Acta Neuropathol. 87. 481〜483 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Itoh.Y: "A variant of Gerstmann-Straussler-Scheinker disease carrying codon 105 mutation with codon 129 polymorphism of the prion proteingene:A clinocopathological study." J Neurol Sci. 127. 77〜86 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Oda.T: "Prion disease with 144 base pair insertion in a Japanese family line." Acta Neuropathol.90. 80〜86 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tateishi.J: "Inherited prion diseases and transmission to rodents." Brain Pathol. 5. 53〜59 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nakagawa.Y: "Apo E in Creutzfeldt-Jakob disease." Lancet. 345. 68 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Furukawa.H: "New variant prion protein in a Japanese family with Gerstmann-Straussler syndrome." Mol.Brain Res.30. 385〜388 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nakagawa.Y: "Apolipoprotein E genotype of sporadic Creutzfeldt-Jakob disease in Japan." Neurosci.Lett.187. 209〜211 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tateishi.J: "First experimental trasmission of fatal familial insomnia." Nature. 376. 434〜435 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hainfellner.JA: "The origanal Gerstmann-Straussler-Scheinker family of Austria;divergent clinico-pathological phenotypes by the constant PrP genotypes." Brain Pathol.5. 201〜211 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Budka H.J: "Tissue handling in suspected Creutfeldt-Jakob disease(CJD)and other human spongiform encephalopathies(prion diseases)." Brain Pathol.5. 319〜322 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Budka.H: "Neuropathological diagnostic criteria for Creutzfeldt-Jakob disease(CJD)and other human spongiform encephalopathies(prion diseases)." Brain Pathol.5. 459〜466 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tateishi.J: "Prion diseases." Microbiol.Immunol.39. 923〜928 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kitamoto.T: "Humanized prion protein knock-in by Cre-inducedsite-specific recombination in the mouse." Biochem.Biophys.Res.Commun.222. 742〜747 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hoque.MZ: "Mutation in the prion protein gene at codon 232 in Japanese patients with Creutzfeldt-Jakob disease;A clinicopathological,immunohistochemical and transmission study." Acta Neuropathol.92. 441〜446 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ghetti.B: "Vascular variant of prionprotein cerebral amyloidosis with tau-positive neurofibrillary tangles:Thephenotype of the stop codon 145 mutation in PRNP." Proc.Natl.Acad.Sci.USA. 93. 744〜748 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tateishi.J: "Experimental transmission of Creutzfeldt-Jakobdisease and related diseases to rodents." Neurology. 46. 532〜537 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kitamoto.T: "Human prion disease and human prion protein disease." Curr.Topics Microbiol.Immunol.207. 27〜34 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Furukawa.H: "A Japapse case of Creutzfeldt-Jakob disease with a point mutation in the prion protein gene at codon 210." J.Neurol.Sci.141. 120〜122 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tateishi.J: "Immunohistochemical evaluation of Creutzfeldt-Jakob disease with reference to the type PrPres deposition." Clin.Neuropathol.15. 358〜360 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nagayama.M: "Fatal familial insomnia with a mutation at codon 178 of the prion protein gene;First report from Japan." Neurology.47. 1313〜1316 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tateishi.J: "Transmission of human prion diseases to rodents." Seminar in Virol 7. 7. 175〜180 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kitamoto T, Nakamura K, Nakao K, Shibuya S, Shin R-W, Gondo Y, Katsuki M, Tateishi J.: "Humanized prion protein knock-in by Cre-inducedsite-specofoc recombination in the mouse." Biochem. Biophys. Res. Commun. 222. 742-747 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hoque MZ, Kitamoto T, Furukawa H, Muramoto T, Tateishi J.: "Mutation inthe prion protein gene at codon 232 in Japanese patients with Creutzfeldt-Jakob disease ; A clinicopathological, immunohistochemical and transmission study." Acta Neuropathol.92. 441-446 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ghetti B, Piccardo P, Spillantini M, Ichimiya Y, Porro M, Perini F, Kitamoto T, Tateishi J, Seilar C, Frangione B, Bugiani O, Giaccone G, Prelli F, Goedert M, Dlouhy SR, Tagliavini F.: "Vascular variant of prionprotein cerebral amyloidosis with tau-positive neurofibrillary tangles : Thephenotype of the stop codon 145 mutation in PRNP." Proc. Natl. Acad. Sci. USA. 93. 744-748 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Tateishi J, Kitamoto T, Hoque MZ, Furukawa H.: "Experimental transmissionof Creutzfeldt-Jakob disease and related diseases to rodents." Neurology. 46. 532-537 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kitamoto T, Tateishi J.: "Human prion disease and human prion pritein disease." Curr. Topics Microbiol. Immunol.207. 27-34 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Furukawa H, Kitamoto T, Hashiguchi H, Tateishi J.: "A Japapse case of Creutzfeldt-Jakob disease with a point mutation in the prion protein geneat codon 210." J. Neurol. Sci. 141. 120-122 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Tateishi J. Kitamoto T, Kretzschmar HA, Mehraein P.: "Immunohistochemical evaluation of Creutzfeldt-Jkob disease with reference to the type PrPres deposition." Chin. Neuropathol. Sci.15. 358-360 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nagayama M, Shinohara Y, Furukawa H, Kitamoto T.: "Fatal familial insomnia with a mutation at codon 178 of the prion protein gene ; First report from Japan." Neurology. 47. 1313-1316 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Tateishi, J.: "Inherited prion disease and transmission to rodents." Brain Pathol.5. 53-59 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nakagawa, Y.: "Allelic variation of apolipoprotein E in Japanese sporadic Creutzfeldt-Jakob disease patients." Neurosic. Lett.187. 209-211 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yamada, T.: "Apolipoprotein E mRNA in the brains of patients with Alzheimer′s disease." J. Neurol. Sci.129. 56-61 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nakagawa, Y.: "Apolipoprotein E in Creutzfeldt-Jakob disease." Lancet. 345. 68- (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Tateishi, J.: "First experimental transmission of fatal familial insomnia." Nature. 376. 434-435 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Furukawa, H.: "New variant prion protein in a Japanese family with Gerstmann-Straussler syndrome." Mol. Brain Res.30. 385-388 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Tateishi, J.: "Prion diseases." Microbiol. Immunol.39. 923-928 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Oda, T.: "Prion disease with 144 base pair insertion in a Japanese family line." Acta Neuropathol.90. 80-86 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kitamoto,T.: "Human prion diseases with bariant prion protein." Phil Trans R Sco Lond B. 343. 391-398 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Inoue,I.: "Japanese family with Creutzfeldt-Jakob disease with cldon 200 point mutation of the prion protein gene." Neurology. 44. 299-301 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yamada.S.: "Creutzfeldt-Jakob disease transmitted by a cadaveric dura matter graft." Neurosurgery. 34. 740-744 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Doi-Yi,R.: "Distribution of prion protein in German patients with Creutzfeldt-Jakob disease is different from that in Japanese patients." Acta Neuropathol. 87. 481-483 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Itoh,Y.: "A variabt of Gerstmann-Straussler-Scheinker disease carrying codon 105 mutation with codon 129 polymorphism of the prion protein gene:A clinicopathological study." J Neurol Sci. 127. 77-86 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Oda,T.: "Prion disease with 144 base pair insertion in a Japanese family line." Acta Neuropathol. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Tateishi,J.: "Inherited prion diseases and transmission to rodents." Brain Pathol. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Nakagawa,Y.: "Apo E in Creutzfeldt-Jakob disease." Lancet. 345. 68 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Furukawa,H.: "New variant prion protein in a Japanese family with Gerstmann-Straussler" Mol Brain Res. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Nakagawa,Y.: "Apolipoprotein E genotype of sporadic Creutzfeldt-Jakob disease in Japan." Neurosci Lett. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Hitotsumatsu,T.: "An exon 8-sliced out transcript of Neurofibromatosis 2 gene is constitutively expressed in various human tissues." J Biochem. 116. 1205-1207 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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