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Knock-out rat production and organ regeneration using embryonic stem cells

Research Project

Project/Area Number 22300147
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory animal science
Research InstitutionNational Institute for Physiological Sciences

Principal Investigator

HIRABAYASHI Masumi  生理学研究所, 行動・代謝分子解析センター, 准教授 (20353435)

Project Period (FY) 2010 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥13,390,000 (Direct Cost: ¥10,300,000、Indirect Cost: ¥3,090,000)
Fiscal Year 2012: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2011: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2010: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Keywords胚性幹細胞 / キメララット / ランダムインテグレーション / ジーンターゲッティング / 臓器欠損 / 4倍体補完 / ノックインラット / ノックアウトラット / 遺伝子ターゲッティング / Pdx1遺伝子 / 相同組換え / エレクトロポレーション / 膵臓欠損 / ラット / トランスジェニック / FGFレセプターインヒビター / MEK活性化インヒビター / GSK3インヒビター / Kusabira-Orange遺伝子 / キメラ
Research Abstract

The ultimate objective of the present study is to produce gene-modified rats useful for regenerative medical research using pluripotent embryonic stem cells (ESCs) and advanced reproductive technologies (ART). Rats expressing the kusabira-orange gene, as well as rats expressing the tdTomato gene knock-in at Rosa26 locus, were successfully produced in preliminary trials. Rats lacking Pdx1 gene were also produced by a similar ART protocol with ESCs (Pdx1-deficient mice have no pancreas), but these rats did not show any phenotypic loss of the pancreas. However, functional rat pancreas was successfully regenerated in adult mice by injecting rat wild-type pluripotent stem cells (PSCs) into Pdx1/KO mouse blastocysts.

Report

(4 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • Research Products

    (50 results)

All 2013 2012 2011 2010 Other

All Journal Article (15 results) (of which Peer Reviewed: 15 results) Presentation (25 results) Book (3 results) Remarks (4 results) Patent(Industrial Property Rights) (3 results) (of which Overseas: 3 results)

  • [Journal Article] A retrospective analysis of germline competence in rat embryonic stem cell Lines2013

    • Author(s)
      Hirabayashi M
    • Journal Title

      Transgenic Res.

      Volume: 22 Issue: 2 Pages: 411-416

    • DOI

      10.1007/s11248-012-9638-7

    • Related Report
      2012 Annual Research Report 2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Identification of rat Rosa26 locus enables generation of knock-in rat lines ubiquitously expressing tdTomato2012

    • Author(s)
      Kobayashi T
    • Journal Title

      Stem Cell

      Volume: Dev. 21 Pages: 2981-2986

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Ability of tetraploid rat blastocysts to support fetal development after complementation with embryonic stem cells2012

    • Author(s)
      Hirabayashi M
    • Journal Title

      Mol. Reprod

      Volume: Dev. 79 Pages: 402-412

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Ability of tetraploid rat blastocysts to support fetal development after complementation with embryonic stem cells2012

    • Author(s)
      Hirabayashi M
    • Journal Title

      Molecular Reproduction and Development

      Volume: 79 Issue: 6 Pages: 402-412

    • DOI

      10.1002/mrd.22043

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Identification of rat Rosa26 locus enables generation of knock-in rat lines ubiquitously expressing tdTomato2012

    • Author(s)
      Kobayashi T
    • Journal Title

      Stem Cell Development

      Volume: 21 Issue: 16 Pages: 2981-2986

    • DOI

      10.1089/scd.2012.0065

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Generation of germline-competent rat induced pluripotent stem cells2011

    • Author(s)
      Hamanaka S
    • Journal Title

      PLoS ONE

      Volume: 6 Issue: 7 Pages: e22008-e22008

    • DOI

      10.1371/journal.pone.0022008

    • Related Report
      2012 Final Research Report 2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Homologous recombination in rat germline stem cells2011

    • Author(s)
      Kanatsu-Shinohara M
    • Journal Title

      Biol. Reprod

      Volume: 85 Pages: 208-217

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Homologous recombination in rat germline stem cells2011

    • Author(s)
      Kanatsu-Shinohara M., et al
    • Journal Title

      Biol Reprod

      Volume: 85 Issue: 1 Pages: 208-17

    • DOI

      10.1095/biolreprod.111.090837

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Embryonic stem (ES) cells and induced-pluripotent stem (iPS) cells in rats2011

    • Author(s)
      Hirabayashi M
    • Journal Title

      Reproductive Medicine & Biology

      Volume: 10 Issue: 4 Pages: 231-238

    • DOI

      10.1007/s12522-011-0095-4

    • NAID

      10030444189

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Generation of rat pancreas in mouse by interspecific blastocyst injection of pluripotent stem cells2010

    • Author(s)
      Kobayashi T
    • Journal Title

      Cell

      Volume: 142 Issue: 5 Pages: 787-799

    • DOI

      10.1016/j.cell.2010.07.039

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Rat transgenesis via embryonic stem cells electroporated with Kusabira-orange gene2010

    • Author(s)
      Hirabayashi M
    • Journal Title

      Mol. Reprod

      Volume: Dev. 77 Issue: 6 Pages: 474-474

    • DOI

      10.1002/mrd.21181

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Impaired active demethylation of paternal genome in pronuclear-stage rat zygotes produced by in vitro fertilization or intracytoplasmic sperm injection.2010

    • Author(s)
      Yoshizawa Y
    • Journal Title

      Mol.Reprod.Dev.

      Volume: 77 Pages: 69-75

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Establishment of rat embryonic stem cell lines that can participate into germline chimerae at high efficiency.2010

    • Author(s)
      Hirabayashi M
    • Journal Title

      Mol.Reprod.Dev.

      Volume: 77 Pages: 94-94

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rat Transgenesis via embryonic stem cells electroporated with kusabira-orange gene.2010

    • Author(s)
      Hirabayashi M
    • Journal Title

      Mol.Reprod.Dev.

      Volume: 77 Pages: 474-474

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Generation of rat pancreas in mouse by interspecific blastocyst injection of pluripotent stem cells.2010

    • Author(s)
      Kobayashi T
    • Journal Title

      Cell

      Volume: 142 Pages: 7897-799

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] マウスおよびラットの多能性幹細胞を用いた異種間4倍体補完法による個体の作製2012

    • Author(s)
      加藤 めぐみ
    • Organizer
      第35回日本分子生物学; Late-breaking Abstracts
    • Place of Presentation
      福岡県福岡市
    • Year and Date
      2012-12-13
    • Related Report
      2012 Final Research Report
  • [Presentation] 免疫不全マウスを用いた異種異所発生によるラット始原生殖細胞からの生殖腺様組織誘導2012

    • Author(s)
      葉山 智工
    • Organizer
      第35回日本分子生物学会
    • Place of Presentation
      福岡県福岡市
    • Year and Date
      2012-12-13
    • Related Report
      2012 Final Research Report
  • [Presentation] ラットES細胞におけるインプリント遺伝子のDNAメチル化解析2012

    • Author(s)
      田村 千尋
    • Organizer
      第105回日本繁殖生物学会
    • Place of Presentation
      茨城県つくば市
    • Year and Date
      2012-09-07
    • Related Report
      2012 Final Research Report
  • [Presentation] Generation of ROSA26- Tomato knock-in rats via gene targeting of pluripotent stem cells2012

    • Author(s)
      Kobayashi T
    • Organizer
      The 10th Annual Meeting of International Society for Stem Cell Research
    • Place of Presentation
      Yokohama, Japan
    • Year and Date
      2012-06-14
    • Related Report
      2012 Final Research Report
  • [Presentation] ラットES細胞株の生殖系列への寄与条件に関する後方視的解析2012

    • Author(s)
      平林 真澄
    • Organizer
      第59回日本実験動物学会
    • Place of Presentation
      大分県別府市
    • Year and Date
      2012-05-26
    • Related Report
      2012 Final Research Report
  • [Presentation] ラットRosa26 遺伝子座の同定と全身性に tdTomato 遺伝子を発現するノックインラットの作製2012

    • Author(s)
      小林 俊寛
    • Organizer
      第59回日本実験動物学会
    • Place of Presentation
      大分県別府市
    • Year and Date
      2012-05-26
    • Related Report
      2012 Final Research Report
  • [Presentation] Generation of Rat Embryo in Mouse by Tetraploid Complementation Using Pluripotent Stem Cells2011

    • Author(s)
      Kobayashi T
    • Organizer
      The 2nid Meeting of World Congress on Reproductive Biology
    • Place of Presentation
      Cairns Convention Centre (Cairns, Australia)
    • Year and Date
      2011-10-10
    • Related Report
      2011 Annual Research Report
  • [Presentation] 4倍体胚盤胞期胚に注入したラットES細胞の産仔発生能について2011

    • Author(s)
      平林真澄
    • Organizer
      第104回日本繁殖生物学会
    • Place of Presentation
      岩手県民情報交流センター(岩手県盛岡市)
    • Year and Date
      2011-09-16
    • Related Report
      2011 Annual Research Report
  • [Presentation] 4倍体胚盤胞期胚に注入したラットES細胞の産仔発生能について2011

    • Author(s)
      平林 真澄
    • Organizer
      第104回日本繁殖生物学会
    • Place of Presentation
      岩手県盛岡市
    • Year and Date
      2011-09-13
    • Related Report
      2012 Final Research Report
  • [Presentation] Generation of Rat Embryo in Mouse by Tetraploid Complementation Using Pluripotent Stem Cells2011

    • Author(s)
      Kobayashi T
    • Organizer
      The 9th Annual Meeting of International Society for Stem Cell Research
    • Place of Presentation
      Toronto, Canada
    • Year and Date
      2011-06-17
    • Related Report
      2012 Final Research Report
  • [Presentation] Generation of germline competent rat induced pluripotent stem cells2011

    • Author(s)
      Hamanaka S
    • Organizer
      The 9th Annual Meeting of International Society for Stem Cell Research
    • Place of Presentation
      Toronto, Canada
    • Year and Date
      2011-06-17
    • Related Report
      2012 Final Research Report
  • [Presentation] 4倍体胚補完によるES/iPS細胞のみに由来するラット産仔作製の試み2011

    • Author(s)
      田村 千尋
    • Organizer
      第58回日本実験動物学会
    • Place of Presentation
      東京都江戸川区
    • Year and Date
      2011-05-25
    • Related Report
      2012 Final Research Report
  • [Presentation] 4倍体胚補完によるES/iPS細胞のみに由来するラット産仔作製の試み2011

    • Author(s)
      田村千尋
    • Organizer
      第58回日本実験動物学会
    • Place of Presentation
      タワーホール船堀(東京都江戸川区)
    • Year and Date
      2011-05-25
    • Related Report
      2011 Annual Research Report
  • [Presentation] Germline transmission可能なラットiPS細胞の樹立2010

    • Author(s)
      濱仲早苗
    • Organizer
      第33回日本分子生物学会
    • Place of Presentation
      兵庫県神戸市
    • Year and Date
      2010-12-10
    • Related Report
      2012 Final Research Report
  • [Presentation] 近交系Brown-Norwayラット胚盤胞に由来するES細胞株の樹立と外来遺伝子導入個体の作製2010

    • Author(s)
      加藤 めぐみ
    • Organizer
      第103回日本繁殖生物学会
    • Place of Presentation
      青森県十和田市
    • Year and Date
      2010-09-03
    • Related Report
      2012 Final Research Report
  • [Presentation] 近交系Brown-Norwayラット胚盤胞に由来するES細胞株の樹立と外来遺伝子導入個体の作製2010

    • Author(s)
      加藤めぐみ
    • Organizer
      第103回日本繁殖生物学会
    • Place of Presentation
      北里大学獣医学部キャンパス(青森県十和田市)
    • Year and Date
      2010-09-03
    • Related Report
      2010 Annual Research Report
  • [Presentation] ラットにおける発生工学の現状と展望2010

    • Author(s)
      平林真澄
    • Organizer
      日本下垂体研究会第25回学術集会
    • Place of Presentation
      伊良湖ガーデンホテル(愛知県田原市)
    • Year and Date
      2010-08-20
    • Related Report
      2010 Annual Research Report
  • [Presentation] ラット胚性幹細胞への外来遺伝子導入がキメラ個体の作製効率に及ぼす影響2010

    • Author(s)
      三宝 誠
    • Organizer
      第57回日本実験動物学会
    • Place of Presentation
      京都府京都市
    • Year and Date
      2010-05-12
    • Related Report
      2012 Final Research Report
  • [Presentation] ラット胚性幹細胞への外来遺伝子導入がキメラ個体の作製効率に及ぼす影響2010

    • Author(s)
      三宝誠
    • Organizer
      第57回日本実験動物学会
    • Place of Presentation
      京都テルサ(京都市南区)
    • Year and Date
      2010-05-12
    • Related Report
      2010 Annual Research Report
  • [Presentation] ラットES細胞株の生殖系列への寄与条件に関する後方視的解析

    • Author(s)
      平林真澄
    • Organizer
      日本実験動物科学・技術 九州2012 第59回日本実験動物学会
    • Place of Presentation
      別府国際コンベンションセンター(大分県)
    • Related Report
      2012 Annual Research Report
  • [Presentation] ラットRosa26 遺伝子座の同定と全身性に tdTomato 遺伝子を発現するノックインラットの作製

    • Author(s)
      小林 俊寛
    • Organizer
      日本実験動物科学・技術 九州2012 第59回日本実験動物学会
    • Place of Presentation
      別府国際コンベンションセンター(大分県)
    • Related Report
      2012 Annual Research Report
  • [Presentation] Generation of ROSA26-Tomato knock-in rats via gene targeting of pluripotent stem cells.

    • Author(s)
      Kobayashi T
    • Organizer
      The 10th Annual Meeting of International Society for Stem Cell Research
    • Place of Presentation
      Pacifico Yokohama(Kanagawa)
    • Related Report
      2012 Annual Research Report
  • [Presentation] ラットES細胞におけるインプリント遺伝子のDNAメチル化解析

    • Author(s)
      田村 千尋
    • Organizer
      第105回日本繁殖生物学会
    • Place of Presentation
      筑波大学 大学会館(茨城県)
    • Related Report
      2012 Annual Research Report
  • [Presentation] 免疫不全マウスを用いた異種異所発生によるラット始原生殖細胞からの生殖腺様組織誘導

    • Author(s)
      葉山 智工
    • Organizer
      第35回日本分子生物学会
    • Place of Presentation
      福岡国際会議場(福岡県)
    • Related Report
      2012 Annual Research Report
  • [Presentation] マウスおよびラットの多能性幹細胞を用いた異種間4倍体補完法による個体の作製

    • Author(s)
      加藤 めぐみ
    • Organizer
      第35回日本分子生物学会
    • Place of Presentation
      福岡国際会議場(福岡県)
    • Related Report
      2012 Annual Research Report
  • [Book] Methodological Advances in the Culture, Manipulation and Utilization of Embryonic Stem Cells for Basic and Practical Applications2011

    • Author(s)
      Hirabayashi M, Hochi S
    • Publisher
      Rat embryonic stem cells: establishment and their use for transgeniesis (Ed. Atwood CA)
    • Related Report
      2012 Final Research Report
  • [Book] Rat embryonic stem cells : establishment and their use for transgenesis "Methodological Advances in the Culture, Manipulation and Utilization of Embryonic Stem Cells for Basic and Practical Applications"((Ed.Atwood CA))2011

    • Author(s)
      Hirabayashi M
    • Total Pages
      13
    • Publisher
      InTech
    • Related Report
      2011 Annual Research Report
  • [Book] Chapter-9 Generation of transgenic rats by ooplasmic injection of sperm cells exposed to exogenous DNA(Rat genomics Methods and Protocols)(Eds.I Anegon)2010

    • Author(s)
      Hirabayashi M
    • Total Pages
      10
    • Publisher
      Humana Press
    • Related Report
      2010 Annual Research Report
  • [Remarks]

    • URL

      http://www.nips.ac.jp/mamtg/

    • Related Report
      2012 Final Research Report
  • [Remarks] 遺伝子改変動物作製室ホームページ

    • URL

      http://www.nips.ac.jp/mamtg/

    • Related Report
      2012 Annual Research Report
  • [Remarks]

    • URL

      http://www.nips.ac.jp/mamtg/

    • Related Report
      2011 Annual Research Report
  • [Remarks]

    • URL

      http://www.nips.ac.jp/mamtg/

    • Related Report
      2010 Annual Research Report
  • [Patent(Industrial Property Rights)] 幹細胞を用いた異種間胚胞キメラ動物作製法 (Method for producing heterologous embryonic chimeric animal using a stem cell)2011

    • Inventor(s)
      中内啓光、小林俊寛、山口智之、濱仲早苗、平林真澄、加藤めぐみ
    • Industrial Property Rights Holder
      国立大学法人東京大学、大学共同利用機関法人自然科学研究機構
    • Filing Date
      2011-07-29
    • Related Report
      2012 Final Research Report
    • Overseas
  • [Patent(Industrial Property Rights)] 抑制性ニューロン特異的にレポーター遺伝子を発現する非ヒト哺乳動物2011

    • Inventor(s)
      柳川右千代、川口泰雄、平林真澄
    • Industrial Property Rights Holder
      国立大学法人群馬大学、大学共同利用機関法人自然科学研究機構
    • Acquisition Date
      2011-12-02
    • Related Report
      2012 Final Research Report
    • Overseas
  • [Patent(Industrial Property Rights)] Method for producing heterogenous embryonic chimeric animal using a stem cell2011

    • Inventor(s)
      Nakauchi H, Kobayashi T, Yamaguchi T, Hamanaka S, Hirabayashi M, Kato M
    • Industrial Property Rights Holder
      Nakauchi H, Kobayashi T, Yamaguchi T, Hamanaka S, Hirabayashi M, Kato M
    • Filing Date
      2011-11-17
    • Related Report
      2011 Annual Research Report
    • Overseas

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Published: 2010-08-23   Modified: 2019-07-29  

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