Generation of human artificial vector to allow the reproduction of gene copy number variation

• Project/Area Number: 20310120
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied genomics
• Research Institution: Tottori University
• Principal Investigator: INOUE Toshiaki 鳥取大学, 医学部, 准教授 (80305573)
• Co-Investigator(Kenkyū-buntansha): KATOH Motonobu 鳥取大学, 医学部, 助教 (00273904) ; OHBAYASHI Tetsuya 鳥取大学, 生命機能研究支援センター, 准教授 (80348804) ; NANBA Eiji 鳥取大学, 生命機能研究支援センター, 教授 (40237631) ; TAKEYA Hiroyuki 崇城大学, 生物生命学部, 教授 (60222105)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2010: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2009: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2008: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
• Keywords: ゲノムコピー数多型 / 人工染色体 / 人工染色体ベクター / コピー数多型

Research Abstract

PhiC31-based recombination allows the multiple introduction of transgenes when the recipient sites(attB sites) are available. We sought to apply this system to the human artificial chromosome(HAC) vector, which can carry the huge size of transgene. We successfully generated the HAC vector carrying multiple copies of attB sites(attB-HAC) in CHO cells. It was confirmed that transgenes tagged with the donor site(attP site) can be loaded onto the attB sites on the attB-HAC, in independent manner of PhiC31 integrase. Intriguingly, Southern blot analyses implicated that multiple copies of transgenes can be loaded onto the attB-HAC in one recombination-reaction in CHO cells. The present study suggests the efficacy of attB-HAC as a tool for reproduction of gene copy number variation in animal models and cell models.

Research Products

• [Journal Article] The STAT3-IGFBP5 axis is critical for IL-6/gp130-induced premature senescence in human fibroblasts (2012)
  • Author(s): Kojima H, Kunimoto H, Inoue T, Nakajima K
  • Journal Title: Cell Cycle Volume: (in press)
  • Peer Reviewed
• [Journal Article] The STAT3-IGFBP5 axis is critical for IL-6/gp 130-induced premature senescence in human fibroblasts (2012)
  • Author(s): 小島裕正
  • Journal Title: Cell Cycle Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] MicroRNA-143 Regulates Human Osteosarcoma Metastasis by Regulating Matrix Metalloprotease-13 Expression (2011)
  • Author(s): Osaki M, Takeshita F, Sugimoto Y, Kosaka N, Yamamoto Y, Yoshioka Y, Kobayashi E, Yamada T, Kawai A, Inoue T, Ito H, Oshimura M, Ochiya T
  • Journal Title: Mol. Ther Volume: 19 Pages: 1123-1130
  • Peer Reviewed
• [Journal Article] Identification of PITX1 as a TERT suppressor gene located on human chromosome 5 (2011)
  • Author(s): Qi DL, Ohhira T, Fujisaki C, Inoue T, Ohta T, Osaki M, Ohshiro E, Seko T, Aoki S, Oshimura M, Kugoh H
  • Journal Title: Mol Cell Biol Volume: 31 Pages: 1624-1636
  • Peer Reviewed
• [Journal Article] SIRT2 downregulation in HeLa can induce p53 accumulation via p38 MAPK activation-dependent p300 decrease, eventually leading to apoptosis (2011)
  • Author(s): Li Y, Matsumori H, Nakayama Y, Osaki M, Kojima H, Kurimasa A, Ito H, Mori S, Katoh M, Oshimura M, Inoue T
  • Journal Title: Genes Cells Volume: 16 Pages: 34-45
  • Peer Reviewed
• [Journal Article] Transcriptional activation of SAP97 by HSF-1 stabilizes Kv1. 5 in HL-1 cells (2011)
  • Author(s): Ting Y, Morikawa K, Kurata Y, Li P, Bahrudin Udin, Miake J, Yamamoto Y, Murata M, Inoue T, Nakai A, Shiota G, Higaki K, Ninomiya H, Shirayoshi, Y, Hisatome I
  • Journal Title: Br J Pharmacol Volume: 162 Pages: 1832-1842
  • Peer Reviewed
• [Journal Article] SIRT2 downregulation in HeLa can induce p53 accumulation via p38 MAPK activation-dependent p300 decrease, eventually leading to apoptosis (2011)
  • Author(s): 李艶沢
  • Journal Title: Genes to Cells Volume: 16 Pages: 34-45
  • Peer Reviewed
• [Journal Article] Transcriptional activation of SAP97 by HSF-1 stabilizes Kv1.5 in HL-1 cells (2011)
  • Author(s): Ya-Kuang Ting
  • Journal Title: British Journal of Pharmacology Volume: 62 Pages: 1832-1842
  • Peer Reviewed
• [Journal Article] Identification of PITX1 as a TERT suppressor gene located on human chromosome 5 (2011)
  • Author(s): 斉東来
  • Journal Title: Molecular and Cellular Biology Volume: 31 Pages: 1624-1636
  • Peer Reviewed
• [Journal Article] MicroRNA-143 Regulates Human Osteosarcoma Metastasis by Regulating Matrix Metalloprotease-13 Expression (2011)
  • Author(s): 尾崎充彦
  • Journal Title: Molecular Therapy Volume: 19 Pages: 1123-1130
  • Peer Reviewed
• [Journal Article] Cysteine-rich protein 61(CYR61) suppresses cell invasion via down-regulation of matrix metalloproteinase-7(MMP-7) expression in a human gastric carcinoma cell line, MKN-45 (2010)
  • Author(s): Osaki M, Inaba A, Nishikawa K, Sugimoto Y, Shomori K, Inoue T, Oshimura M, Ito H
  • Journal Title: Mol Med Rep Volume: 3 Pages: 711-715
  • Peer Reviewed
• [Journal Article] Cysteine-rich protein 61 (CYR61) suppresses cell invasion via down-regulation of matrix metalloproteinase-7 (MMP-7) expression in a human gastric carcinoma cell line, MKN-45 (2010)
  • Author(s): 尾崎充彦
  • Journal Title: Molecular Medicine Report Volume: 3 Pages: 711-715
  • Peer Reviewed
• [Journal Article] SIRT2 downregulation confers resistance to microtubule inhibitors by prolonging chronic mitotic arrest (2009)
  • Author(s): Inoue T, Nakayama Y, Yamada H, Li YC, Yamaguchi S, Osaki M, Kurimasa A, Hiratsuka M, Katoh M, Oshimura M
  • Journal Title: Cell Cycle Volume: 8 Pages: 1279-1291
  • Peer Reviewed
• [Journal Article] Enhanced apoptosis during early neuronal differentiation in mouse ES cells with autosomal imbalance (2009)
  • Author(s): Kai Y, Wang CC, Kishigami S, Kazuki Y, Abe S, Takiguchi M, Shirayoshi Y, Inoue T, Ito H, Wakayama T, Oshimura M
  • Journal Title: Cell Res Volume: 19 Pages: 247-258
  • Peer Reviewed
• [Journal Article] SIRT2 downregulation confers resistance to microtubule inhibitors by prolonging chronic mitotic arrest. (2009)
  • Author(s): 井上敏昭
  • Journal Title: Cell Cycle 8 Pages: 1279-1291
  • Peer Reviewed
• [Journal Article] Enhanced apoptosis during early neuronal differentiationin mouse ES cells with autosomal imbalance. (2009)
  • Author(s): Kai Y.
  • Journal Title: Cell Ees. 19 Pages: 247-258
  • Peer Reviewed
• [Journal Article] SIRT2 downregulation confers resistance to microtubuleinhibitors by prolonging chronic mitotic arrest (2009)
  • Author(s): Inoue T.
  • Journal Title: Cell Cycle 8 Pages: 1279-1291
  • Peer Reviewed
• [Journal Article] Functional stabilization of Kv1. 5 protein by Hsp70 in mammalian cell lines (2008)
  • Author(s): Hirota Y, Kurata Y, Kato M, Notsu T, Koshida S, Inoue T, Kawata Y, Miake J, Bahrudin U, Li P, Hoshikawa Y, Yamamoto Y, Igawa O, Shirayoshi Y, Nakai A, Ninomiya H, Higaki K, Hiraoka M, Hisatome I
  • Journal Title: Biochem Biophys Res Commun Volume: 372 Pages: 469-474
  • Peer Reviewed
• [Journal Article] Introduction of a CD40L genomic fragment via a human artificial chromosome vector permits cell-type-specific gene expression and induces immunoglobulin secretion (2008)
  • Author(s): Yamada H, Li YC, Nishikawa M, Oshimura M, Inoue T
  • Journal Title: J Hum Genet Volume: 53 Pages: 447-453
  • NAID: 10021248965
  • Peer Reviewed
• [Journal Article] Introduction of a CD40L genomic fragment via a human artificial chromosome vector permits cell-type-specific gene expression and induces immunoglobulin secretion. (2008)
  • Author(s): Yamada H.
  • Journal Title: J. Hum Genet. 53 Pages: 447-453
  • Peer Reviewed
• [Journal Article] Functional stabilization of Kv1.5 protein by Hsp70 in mammalian cell lines. (2008)
  • Author(s): Hirota Y.
  • Journal Title: Biochem Biophys Res Commun. 372 Pages: 469-474
  • Peer Reviewed
• [Journal Article] Cysteine-rich protein 61 (CYR61) suppresses cell invasion via down-regulation of matrix metalloproteinase-7 (MMP-7) expression in a human gastric carcinoma cell line, MKN-45
  • Author(s): 尾崎充彦
  • Journal Title: Molecular Medicine Report (印刷中)
  • Peer Reviewed
• [Presentation] 複数の遺伝子搭載が可能な人工染色体ベクターの構築 (2011)
  • Author(s): 松森はるか、加藤基伸、中山祐二、押村光雄、井上敏昭
  • Organizer: 日本農芸化学会2011年度大会
  • Place of Presentation: 京都会館(京都市)
  • Year and Date: 2011-03-25
• [Presentation] 麻疹ウイルスエンベロープを利用した微小核細胞融合法による染色体導入 (2011)
  • Author(s): 高橋悠、加藤基伸、香月康宏、香月加奈子、梶谷尚世、滝口正人、中山祐二、中村貴史、井上敏昭、押村光雄
  • Organizer: 日本農芸化学会2011年度大会
  • Place of Presentation: 京都会館(京都市)
  • Year and Date: 2011-03-25
• [Presentation] 複数の遺伝子搭載が可能な人工染色体ベクターの構築 (2011)
  • Author(s): 松森はるか
  • Organizer: 日本農芸化学会2011年度大会ただし東日本大震災の影響で学会中止、誌上発表のみ
  • Place of Presentation: 京都会館(京都市)
  • Year and Date: 2011-03-25
• [Presentation] 麻疹ウイルスエンベロープを利用した微小核細胞融合法による染色体導入 (2011)
  • Author(s): 高橋悠
  • Organizer: 日本農芸化学会2011年度大会ただし東日本大震災の影響で学会中止、誌上発表のみ
  • Place of Presentation: 京都会館(京都市)
  • Year and Date: 2011-03-25
• [Presentation] 複数の遺伝子搭載が可能な人工染色体ベクターの構築 (2011)
  • Author(s): 荻野由加利、松森はるか、押村光雄、中山祐二、井上敏昭
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
• [Presentation] がん治療の新たな標的分子としてのSIRT2 (2011)
  • Author(s): 李艶沢、末松知久、中山祐二、押村光雄、井上敏昭
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
• [Presentation] 複数の遺伝子搭載が可能な人工染色体ベクターの構築 (2011)
  • Author(s): 荻野由加利
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)(ポスター発表)(口頭発表)
• [Presentation] がん治療の新たな標的分子としてのSIRT2 (2011)
  • Author(s): 李艶沢
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)(口頭発表)(ポスター発表)
• [Presentation] Chromosome transfer via microcell fusion utilizing fusogenic envelope proteins of Measles virus (2010)
  • Author(s): Motonobu Katoh, Yasihiro Kazuki, Haruka Takahashi, Kanako Kazuki, Naoyo Kajitani, Masato Takiguchi, Yuji Nakayama, Takafumi Nakamura, Toshiaki Inoue, Mitsuo Oshimura
  • Organizer: 第33回日本分子生物学会第83回日本生化学会合同大会
  • Place of Presentation: 神戸ポートピアアイランド(神戸市)
  • Year and Date: 2010-12-10
• [Presentation] SIRT2発現低下によるp53依存性細胞死誘導とその機序 (2010)
  • Author(s): 李艶沢、井上敏昭、押村光雄
  • Organizer: 第69回日本癌学会学術総会
  • Place of Presentation: 大阪国際会議場(大阪市)
  • Year and Date: 2010-09-23
• [Presentation] ヒト5番染色体上に存在する新規テロメレース抑制遺伝子の同定 (2010)
  • Author(s): 久郷裕之、Dong-Lai Qi、大平嵩人、太田力、井上敏昭、押村光雄
  • Organizer: 第69回日本癌学会学術総会
  • Place of Presentation: 大阪国際会議場(大阪市)
  • Year and Date: 2010-09-22
• [Presentation] SIRT2発現低下による微小管阻害剤耐性獲得とその分子機構 (2009)
  • Author(s): 李艶沢、押村光雄、井上敏昭
  • Organizer: 第68回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 2009-10-03
• [Presentation] 人工染色体システムを用いたトリプレットリピート解析系の構築 (2008)
  • Author(s): 中山祐二、井上敏昭、押村光雄、難波栄二
  • Organizer: 第53回日本人類遺伝学会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 2008-09-27
• [Book] 実験医学別冊「実験医学別冊培養細胞実験ハンドブック」細胞融合法 (2009)
  • Author(s): 則兼聡子、井上敏昭、押村光雄
  • Publisher: 羊土社
• [Book] 実験医学別冊「実験医学別冊改訂培養細胞実験ハンドブック」細胞融合法 (2008)
  • Author(s): 則兼聡子
  • Publisher: 羊土社
• [Remarks] 鳥取大学医学部生命科学科ゲノム医工学分野
  • URL: http://www.med.tottori-u.ac.jp/genome/309/
• [Remarks] 鳥取大学染色体工学研究センター
  • URL: http://www.med.tottori-u.ac.jp/chromosome/
• [Remarks] 鳥取大学医学部生命科学科ゲノム医工学分野
  • URL: http://www.med.tottori-u.ac.jp/genome/309/
• [Remarks] 鳥取大学染色体工学研究センター
  • URL: http://www.med.tottori-u.ac.jp/chromosome/
• [Remarks]
  • URL: http://www.med.tottori-u.ac.jp/p/igaku/gakka/seimei/bunshi/genome_home/genome_home1/
• [Remarks]
  • URL: http://www.med.tottori-u.ac.jp/p/igaku/daigakuin/seimei/genome_home/