Improvement of high-throughput method to generate the knock-in ES

• Project/Area Number: 20310123
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied genomics
• Research Institution: Kazusa DNA Research Institute
• Principal Investigator: NAKAYAMA Manabu Kazusa DNA Research Institute, ヒトゲノム研究部・ヒト遺伝子応用技術研究室, 主任研究員 (30370927)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥16,900,000 (Direct Cost: ¥13,000,000、Indirect Cost: ¥3,900,000); Fiscal Year 2010: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2009: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2008: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
• Keywords: 遺伝子 / ゲノム / 機能ゲノム / 動物 / 発生・分化 / バイオテクノロジー / 病態モデル動物

Research Abstract

Producing of knock-in mouse is extremely useful method to analyze various proteins' function in mice. By using Red/ET recombination system which can be recombined any DNA sequence through homologous recombination reaction, we have developed a high-through system to construct targeting vectors for knock-in and a new system to introduce exchangeable protein tag to C-terminal end of the protein. We also have developed two new site-specific recombination systems to engineer genome of mouse ES cells.

Research Products

• [Journal Article] Recombinase-mediated cassette exchange (RMCE) and BAC engineering via VCre/VloxP and SCre/SloxP systems. (2011)
  • Author(s): Minorikawa S., Nakayama M.
  • Journal Title: BioTechniques Vol.50 Pages: 235-246
  • Peer Reviewed
• [Journal Article] VCre/VloxP and SCre/SloxP : new site-specific recombination systems for genome engineering. (2011)
  • Author(s): Suzuki E., Nakayama M.
  • Journal Title: Nucleic Acids Res. Vol.39
  • Peer Reviewed
• [Journal Article] VCre/VloxP and SCre/SloxP : new site-specific recombination systems for genome engineering (2011)
  • Author(s): Suzuki E.
  • Journal Title: Nucleic Acids Res. Volume: 39
  • Peer Reviewed
• [Journal Article] Recombinase-mediated cassette exchange (RMCE) and BAC engineering via VCre/VloxP and SCre/SloxP systems (2011)
  • Author(s): Minorikawa S.
  • Journal Title: BioTechniques Volume: 50 Pages: 235-246
  • Peer Reviewed
• [Journal Article] Extracellular Calcium Controls Background Current and Neuronal Excitability via an UNC79-UNC80-NALCN Cation Channel Complex. (2010)
  • Author(s): Lu B., Zhang Q., Wang H., Wang Y., Nakayama M., Ren D.
  • Journal Title: Neuron Vol.68 Pages: 488-499
  • Peer Reviewed
• [Journal Article] Defining the immunological phenotype of Fc receptor-like B (FcRLB) deficient mice : confounding role of the inhibitory Fc γ RIIb. (2010)
  • Author(s): Masuda K., Mori H., Ohara O., Nakayama M., Wang J., Burrows P.
  • Journal Title: Cellular Immunology Vol.266 Pages: 24-31
  • Peer Reviewed
• [Journal Article] Homologous recombination in human iPS and ES cells for use in gene correction therapy. (2010)
  • Author(s): Nakayama M.
  • Journal Title: Drug Discov.Today Vol.15 Pages: 198-202
  • Peer Reviewed
• [Journal Article] Eliminating target cells by inducing apoptosis-related factors using hTERT promoter. (2010)
  • Author(s): Takashina T., Nakayama M.
  • Journal Title: Open Biotechnol J Vol.4 Pages: 1-7
  • Peer Reviewed
• [Journal Article] Extracellular Calcium Controls Background Current and Neuronal Excit ability via an UNC79-UNC80-NALCN Cation Channel Complex. (2010)
  • Author(s): Lu B.
  • Journal Title: Neuron. Volume: 68 Pages: 488-499
  • Peer Reviewed
• [Journal Article] Homologous recombination in human iPS and ES cells for use in gene correction therapy (2010)
  • Author(s): Nakayama M.
  • Journal Title: Drug Discov.Today 15 Pages: 198-202
  • Peer Reviewed
• [Journal Article] Eliminating target cells by inducing apoptosis-related factors using hTERT promoter (2010)
  • Author(s): Takashina T.
  • Journal Title: Open Biotechnol J 4 Pages: 1-7
  • Peer Reviewed
• [Journal Article] Requirement of MARCH-I-mediated MHC II ubiquitination for the maintenance of conventional dendritic cells. (2009)
  • Author(s): Ohmura-Hoshino M, Matsuki Y, Mito-Yoshida M, Goto E, Aoki-Kawasumi M, Nakayama M, Ohara O, Ishido S
  • Journal Title: J Immunol. Vol.11 Pages: 6893-6897
  • Peer Reviewed
• [Journal Article] Engineered FADD induces apoptosis via an artificial death-inducing signaling complex (DISC). (2009)
  • Author(s): Suzuki E, Takashina T., Nakayama M.
  • Journal Title: Int J Biomed Sci. Vol.5 Pages: 237-245
  • Peer Reviewed
• [Journal Article] Cell therapy using induced pluripotent stem (iPS) cells meets next-next generation DNA sequencing technology. (2009)
  • Author(s): Nakayama M.
  • Journal Title: Curr.Genomics Vol.10 Pages: 303-305
  • Peer Reviewed
• [Journal Article] Engineered FADD induces apoptosis via an artificial death-inducing signaling complex(DISC) (2009)
  • Author(s): Suzuki E.
  • Journal Title: Int J Biomed Sci. 5 Pages: 237-245
  • Peer Reviewed
• [Journal Article] Cell therapy using induced pluripotent stem(iPS)cells meets next-next generation DNA sequencing technology (2009)
  • Author(s): Nakayama M.
  • Journal Title: Current Genomics 10 Pages: 303-305
  • Peer Reviewed
• [Journal Article] The zinc transporter SLC39A13/ZIP13 is required for connective tissue development ; its involvement in BMP/TGF-beta signaling pathways. (2008)
  • Author(s): Fukada T, Civic N, Furuichi T, Shimoda S, Mishima K, Higashiyama H, Idaira Y, Asada Y, Kitamura H, Yamasaki S, Hojyo S, Nakayama M, Ohara O, Koseki H, Dos Santos HG, Bonafe L, Ha-Vinh R, Zankl A, Unger S, Kraenzlin ME, Beckmann JS, Saito I, Rivolta C, Ikegawa S, Superti-Furga A, Hirano T
  • Journal Title: PLoS ONE. Vol.3
  • Peer Reviewed
• [Journal Article] Cell therapy using induced pluripotent stem (iPS) cells meets next-next generation DNA sequencing technology
  • Author(s): M. Nakayama
  • Journal Title: Current Genomics (In press)
  • Peer Reviewed
• [Presentation] 新規部位特異的組み換えシステムを用いたゲノム改変技術の開発 (2010)
  • Author(s): 中山学
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 神戸ポートピアホテル
  • Year and Date: 2010-12-07
• [Presentation] ノックイン胚性幹細胞作製の迅速化と新規ゲノム改変技術の開発 (2009)
  • Author(s): 中山学
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2009-12-11
• [Presentation] 包括的機能研究のためのノックイン胚性幹細胞作製の迅速化と解析 (2008)
  • Author(s): 中山学
  • Organizer: BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会合同大会)
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2008-12-11
• [Presentation] 包括的機能研究のためのノックイン胚性幹細胞作製の迅速化と解析 (2008)
  • Author(s): 中山学
  • Organizer: BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会)
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2008-12-11
• [Remarks] ホームページ等
• [Patent(Industrial Property Rights)] 新規な部位特異的組換え酵素とその認識配列を用いた部位特異的組換え方法 (2009)
  • Inventor(s): 中山学
  • Industrial Property Rights Holder: かずさDNA研究所
  • Filing Date: 2009-10-29
• [Patent(Industrial Property Rights)] 新規な部位特異的組換え酵素とその認識配列を用いた部位特異的組換え方法 (2009)
  • Inventor(s): 中山学
  • Industrial Property Rights Holder: かずさDNA研究所
  • Industrial Property Number: 2009-136830
  • Filing Date: 2009-06-08