Application of a novel gene-disruption technique based on the suppression of NMD for the T-lymphocyte research

• Project/Area Number: 21310128
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied genomics
• Research Institution: Nara Institute of Science and Technology
• Principal Investigator: YASUMASA Ishida 奈良先端科学技術大学院大学, バイオサイエンス研究科, 准教授 (10221756)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥16,510,000 (Direct Cost: ¥12,700,000、Indirect Cost: ¥3,810,000); Fiscal Year 2011: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2010: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2009: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: 遺伝子トラップ / マウス / ES細胞 / nonsense-mediated mRNA decay / リンパ球 / マウスES細胞 / ノックアウトマウス / Tリンパ球 / UPATrap / 遺伝子発現

Research Abstract

In order to improve the efficiency of random insertional mutagenesis, the backbone of an NMD-suppressing poly(A)-trap technique "UPATrap" has been changed from a standard retrovirus to a DNA transposon Tol2. This modification increased the disruption rate of transcriptionally silent genes in mouse embryonic stem(ES) cells and permitted conditional inactivation of trapped genes. In addition, we solved a long-standing problem associated with the transposon vector system by incorporating a mixture of 15 differentially tagged Tol2 transposons. These developments will certainly accelerate the analyses on the nature of T lymphocytes.

Research Products

• [Journal Article] CtBP-interacting BTB zinc finger protein(CIBZ) promotes proliferation and G1/S transition in embryonic stem cells via Nanog. (2012)
  • Author(s): Nishii, T., Oikawa, Y., Ishida, Y., Kawaichi, M., and Matsuda, E.
  • Journal Title: J. Biol. Chem Volume: Vol.287 Pages: 12417-12424
  • NAID: 120005818445
  • Peer Reviewed
• [Journal Article] Mixture of differentially tagged Tol2 transposons accelerates conditional disruption of a broad spectrum of genes in mouse embryonic stem cells (2012)
  • Author(s): N.I.Mayasari, et al
  • Journal Title: Nucleic Acids Research Volume: 40(In Press) Issue: 13 Pages: e97-e97
  • DOI: 10.1093/nar/gks262
  • Peer Reviewed
• [Journal Article] The methyl-CpG-binding protein CIBZ suppresses myogenic differentiation by directly inhibiting myogenin expression (2011)
  • Author(s): Oikawa, Y., Omori, R., Nishii, T., Ishida, Y., Kawaichi, M., and Matsuda, E.
  • Journal Title: Cell Res Volume: Vol.21 Pages: 1578-1590
  • NAID: 120005818451
  • Peer Reviewed
• [Journal Article] Evidence that the Upf1-related molecular motor scans the 3'-UTR to ensure mRNA integrity.
  • Author(s): Shigeoka, T., Kato, S., Kawaichi, M., and Ishida, Y.
  • Journal Title: Nucleic Acids Res Volume: (in press)
  • Peer Reviewed
• [Journal Article] Mixture of differentially tagged Tol2 transposons accelerates conditional disruption of a broad spectrum of genes in mouse embryonic stem cells.
  • Author(s): Mayasari, N. I., Mukougawa, K., Shigeoka, T., Kawakami, K., Kawaichi, M, and Ishida, Y.
  • Journal Title: Nucleic Acids Res Volume: (in press)
  • Peer Reviewed
• [Presentation] Mixture of differentially tagged Tol2 transposons accelerates conditional disruption of a broad spectrum of genes in mouse embryonic stem cells. (2012)
  • Author(s): Yasumasa Ishida(石田靖雅)
  • Organizer: The Company of Biologists Workshop "New Technologies and Applications for Genome Engineering"
  • Place of Presentation: Steiyning, UK(英国)
  • Year and Date: 2012-03-25
• [Presentation] Mixture of differentially tagged Tol2 transposons accelerates conditional disruption of a broad spectrum of genes in mouse embryonic stem cells (2012)
  • Author(s): 石田靖雅
  • Organizer: The Company of Biologists Workshop "New Technologies and Applications for Genome Engineering"
  • Place of Presentation: Wiston House, Steyning, UK
  • Year and Date: 2012-03-25
• [Presentation] 条件的遺伝子改変ES細胞株の量産とデータベース化 (2011)
  • Author(s): N. I. Mayasari、田畑海渡、石田靖雅
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 条件的遺伝子改変ES細胞株の量産とデータベース化 (2011)
  • Author(s): N.I.Mayasari
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(複数日ポスター発表)
• [Remarks] The NAISTrap project
  • URL: http://bsw3.naist.jp/kawaichi/naistrap.html
• [Remarks] The NAISTrap project
  • URL: http://bsw3.naist.jp/kawaichi/naistrap.html