Development of highly effective gene knock-in procedure using neural crest-specific gene expression machinery

• Project/Area Number: 22700449
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Laboratory animal science
• Research Institution: Kyoto University
• Principal Investigator: SATO Takahiro 京都大学, iPS細胞研究所, 特任研究員 (10572000)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2010: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 発生工学

Research Abstract

To identify the neural crest cell-specific enhancer region as well as transcription factors that bind to the enhancer region, we conducted luciferase assay using several upstream promoter regions on endothelin type-A receptor gene locus. Consequently, we found that about 30 bases as a critical enhancer sequence that consisted of presumable tandem binding sites of two transcription factors. In vivo experiments in transgenic mice exhibited an insufficiency of this enhancer sequence for neural crest cell-specific forced expression of LacZ gene. These results suggest that a possible coordination with other region may be required to proper gene expression in neural crest cells.
Adenoviral transduction in fertilized eggs exhibited toxicity for their later normal development, whereas strong expression of LacZ was observed. Therefore, the important issue of viral toxicity remained to be overcome.

Research Products

• [Journal Article] Identification and developmental analysis of endothelin type-A expressing cells in the mouse kidney (2011)
  • Author(s): Kitazawa T, Sato T, Nishiyama K, Asai R, Arima Y, Uchijima Y, Kurihara Y, Kurihara H
  • Journal Title: Gene Expr Patterns Volume: 11 Pages: 371-7
  • Peer Reviewed
• [Journal Article] Identification and developmental analysis of endothelin receptor type-A expressing cells in the mouse kidney (2011)
  • Author(s): Kitazawa T, Sato T, Nishiyama K, Asai R, Arima Y, Uchijima Y, Kurihara Y, Kurihara H.
  • Journal Title: Gene Expr. Patterns Volume: 11 Issue: 7 Pages: 371-377
  • DOI: 10.1016/j.gep.2011.04.001
  • Peer Reviewed
• [Journal Article] Endothelin receptor type-A expression defines a distinct cardiac subdomain within the heart field, with a later implication of this signaling pathway in chamber myocardium formation (2010)
  • Author(s): Asai R, Kurihara Y, Fujisawa K, Sato T, Kawamura Y, Kokubo H, Tonami K, Nishiyama K, Uchijima Y, Miyagawa-Tomita S, Kurihara H.
  • Journal Title: Development Volume: 137 Issue: 22 Pages: 3823-3833
  • DOI: 10.1242/dev.054015
  • Peer Reviewed
• [Journal Article] Endothelin receptor type A expression defines a distinctcardiac subdomain within the heart field and is later implicated in chamber myocardium formation. (2010)
  • Author(s): Asai R, Kurihara Y, Fujisawa K, Sato T, Kawamura Y, Kokubo H, Tonami K, Nishiyama K, Uchijima Y, Miyagawa-Tomita S, Kurihara H.
  • Journal Title: Development Volume: 137 Pages: 3823-3833
  • Peer Reviewed
• [Presentation] Hox gene Function and possible crosstalk with Dlx genes in cranofacial morphogenesis (2011)
  • Author(s): Kitazawa T, Fujisawa K, Kurihara Y, Arima Y, Kawamura Y, Sato T, Uchijima Y, Kurihara H
  • Organizer: The 34th Annual Meeting of the Molecular Biology Society of Japan
  • Place of Presentation: Yohohama
  • Year and Date: 2011-12-13
• [Presentation] Hox gene Function and possible crosstalk with Dlx genes in cranofacial morphogenesis (2011)
  • Author(s): Kitazawa T, Fujisawa K, Kurihara Y, Arima Y, Kawamura Y, Sato T, Uchijima Y, Kurihara H
  • Organizer: The 34th Annual Meeting of the Molecular Biology Society of Japan
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2011-12-13
• [Presentation] Endothelin-1/Endothelin type-A receptor signaling regulates pharyngeal arch artery development through Dlx5/6-independent pathway (2011)
  • Author(s): Kim KS, Arima Y, Asai R, Sato T, Nishiyama K, Kurihara H
  • Organizer: The 19th Annual Meeting of the Japanese Vascular Biology and Medicine Organization/The 1st Asia-Pscific Vascular Biology Meeting
  • Place of Presentation: Tokyo
  • Year and Date: 2011-12-10
• [Presentation] Endothelin-1/Endothelin type-A receptor signaling regulates pharyngeal arch artery development through Dlx5/6-independent pathway (2011)
  • Author(s): Kim KS, Arima Y, Asai R, Sato T, Nishiyama K, Kurihara H
  • Organizer: The 19th Annual Meeting of the Japanese Vascular Biology and Medicine Organization The 1st Asia-Pscific Vascular Biology Meeting
  • Place of Presentation: Tokyo Station Conference
  • Year and Date: 2011-12-10
• [Presentation] Endothelin type-A receptor expression defines a distinct subpoplation within the first heart field contributing to chamber myocardium (2010)
  • Author(s): Asai R, Kurihara Y, Fujisawa K, Sato T, Kawamura Y, Kokubo H, Tonami K, Nishiyama K, Uchijima Y, Miyagawa-Tomita S, Kurihara H
  • Organizer: BMB2010
  • Place of Presentation: Kobe
  • Year and Date: 2010-12-10
• [Presentation] Endothelin type-A receptor expression defines a distinct subpoplation within the first heart field contributing to chamber myocardium (2010)
  • Author(s): Asai R, Kurihara Y, Fujisawa K, Sato T, Kawamura Y, Kokubo H, Tonami K, Nishiyama K, Uchijima Y, Miyagawa-Tomita S, Kurihara H.
  • Organizer: BMB2010
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-10
• [Presentation] Involvment of non-coding RNA in the Endothelin signaling in branchal arch development (2010)
  • Author(s): Kurihara Y, Uchijima Y, Fujisawa K, Sato T, Kushiyama S, Asai R, Kitazawa T, Nishiyama K, Kurihara H
  • Organizer: BMB2010
  • Place of Presentation: Kobe
  • Year and Date: 2010-12-08
• [Presentation] Transcription regulation of non-coding RNA Evf-2 in the Endothelin-1/Dlx5/Dlx6 pathway (2010)
  • Author(s): Uchijima Y, Kurihara Y, Sato T, Fujisawa K, Kushiyama S, Kurihara H
  • Organizer: BMB2010
  • Place of Presentation: Kobe
  • Year and Date: 2010-12-08
• [Presentation] Transcription regulation of non-coding RNA Evf-2 in the Endothelin-1/Dlx5/Dlx6 pathway (2010)
  • Author(s): Uchijima Y, Kurihara Y, Sato T, Fujisawa K, Kushiyama S, Kurihara H.
  • Organizer: BMB2010
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-08
• [Presentation] Involvment of non-coding RNA in the Endothelin signaling in branchal arch development (2010)
  • Author(s): Kurihara Y, Uchijima Y, Fujisawa K, Sato T, Kushiyama S, Asai R, Kitazawa T, Nishiyama K, Kurihara H.
  • Organizer: BMB2010
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-08
• [Presentation] Involvement of the endothelin-1/endothelin receptor type-A signaling in determination of the mandibular identity in mouse craniofacial development (2010)
  • Author(s): Sato T, Kurihara Y, Asai R, Kawamura Y, Tonami K, Uchijima Y, Heude E, Ekker M, Levi G, Kurihara H
  • Organizer: Gordon Research Conrference : Craniofacial Morphogenesis & Tissue Regeneration
  • Place of Presentation: Lucca(Barga), Italy
• [Presentation] Non-coding RNA regulation of endothelin signaling in branchal arch development (2010)
  • Author(s): Kurihara Y, Uchijima Y, Fujisawa K, Sato T, Kushiyama S, Asai R, Kitazawa T, Nishiyama K, Kurihara H.
  • Organizer: Gordon Research Conrference : Craniofacial Morphogenesis & Tissue Regeneration
  • Place of Presentation: Lucca(Barga), Italy
• [Presentation] Involvement of the endothelin-1/endothelin receptortype-A signaling in determination of the mandibular identity in mouse craniofacial development (2010)
  • Author(s): Sato T, Kurihara Y, Asai R, Kawamura Y, Tonami K, UchijimaY, Heude E, Ekker M, Levi G, Kurihara H.
  • Organizer: Gordon Research Conrference : Craniofacial Morphogenesis & Tissue Regeneration
  • Place of Presentation: Lucca (Barga), Italy
• [Presentation] Non-coding RNA regulation of endothelin signaling in branchal arch development (2010)
  • Author(s): Kurihara Y, Uchijima Y, Fujisawa K, Sato T, Kushiyama S, Asai R, Kitazawa T, Nishiyama K, Kurihara H.
  • Organizer: Gordon Research Conrference : Craniofacial Morphogenesis & Tissue Regeneration
  • Place of Presentation: Lucca (Barga), Italy
• [Remarks]
  • URL: http://www.cira.kyoto-u.ac.jp/i/index.html