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Precise Manipulation of Developemental transgenic cells using a microwichirned Technology

Research Project

Project/Area Number 05455005
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 広領域
Research InstitutionJapan Advanced Institute of Science & Technology (JAIST)

Principal Investigator

TAMIYA Eiichi  JAIST,Materials Science Professor, 材料科学研究所, 教授 (60179893)

Co-Investigator(Kenkyū-buntansha) YOKOYAMA Kenji  JAIST,Materials Science Assoc. Prof., 材料科学研究所, 助教授 (80242121)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥7,500,000 (Direct Cost: ¥7,500,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1993: ¥6,000,000 (Direct Cost: ¥6,000,000)
KeywordsMicromachinning technique / Embryo Engineering / Medaka / Gene transfer / Electroporation / Transgluic fish / トランスジェニック生物 / 半導体集積化技術 / 遺伝子導入法 / エレクトロポーレション / 微小電極
Research Abstract

In this study, a novel system for gene transfer has been fabricated using micro-machining techniques. Micromachining techniques have been developed for fabricating semiconductor devices. Microelectrodes and can be fabricated onto the glass plate using lithography and vapor deposition techniques.
We have used Medaka (Oryzias latipes) as an experimental fish, because it is a suitable material for embryo engineering research based on the following reasons. Compared with mice, fish fertilizes eggs extemally, and there is no need for implant eggs into the uterus of pseudo pregnant animal. Since fish has a transparent egg membrane, both developement and differentiation process are easy to observe.
A high electric field induces a hole of lipid bilayr of the cell membranes and results in destroying the egg yolk and a limited yield. Therefore, the electric field should be localized to a certain part of cytoplasm of the cell to increase the yield. Micro-electrodes were fabricated on glass plate with micro-machining techniques to introduce gene to them. The material of the bottom of the electrode is Cr, and the upper part is Au. A polymer film with a hole (1.2mm in the diameter) is attached with an adhesive.
Microelectrodes was used for application of localized high electric field at the animal pole of the fertilized egg. High electric field pulses (750 V/cm-1000V/cm) of duration fifty microseconds was applied at a repetition rate of 1Hz for five seconds. Microscopic observation showed no damage of eggs using our system.
Forty-one percent of the eggs show luminescence (19 samples in 46 samples). The present system gave a higher ratio of the expression of luciferase than a conventional electroporation method did.
The dependence on the cleavage stages about electroporation was measured.
Gene transfer was observed in the range from 1-cell stage to 64-cell stage embryo.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Y.Murakami: "Micromachined electroporation system for transglnic fish" J.Biotechnology. 34. 35-42 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 民谷栄一: "微小薄膜電極を用いる生物分子・細胞の操作" 化学. 49. 746-747 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 民谷栄一: "マイクロバイオセンサーの開発動向と展望" 計測と制御. 134. 36-41 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Murakami: "Micromachied Electrochemical Flow Ceel for Bioseusing" Electroavalysis. 6. 735-739 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] E.Tamiya: "Direct Monitoring of Glutamate Deleased from-" Sensors & Materials. 7(in press). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 民谷栄一: "バイオセンサーによる細胞情報の解析" 農林水産技術「研究ジャーナル」. 18. 29-33 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 民谷栄一: "機械分析ガイドブック" 丸善(印刷中), (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] E.Tamiya: "Olfaction and TasteXI" Springer-velag, (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Murakami et al.: "Micromachined electroporation system for transgluic fish." J.Biotechnology. 34. 35-42 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Murakami et al.: "Micromachined Electrochemical Flow Cell for Biosensing." Electroanalysis. 6. 735-739 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] E.Tamiya et al.: "Direct Monitoring of Glutamate Released from Cerekellum neurous" Sensors & Materials. 7(in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Murakami: "Micromachined electroporation system for fransgluic fish" J.Biotechnlogy. 34. 35-42 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 民谷栄一: "微小薄膜電極を用いる生体分子・細胞の操作" 化学. 49. 746-747 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 民谷栄一: "マイクロバイオセンサの開発動向と展望" 計測と制御. 134. 36-41 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] E.Tamiya: "Direct Monitoring of Glutamate Released from Cerebellar NeurcnS Vsingo." Sensors & Mcteials. 7(in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Y.Murakami: "Micromachined Dlectrochemical Flom Cell for Biosensing" Electroonalysis. 6. 735-739 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 民谷栄一: "バイオセンサーによる細胞情報の解析" 農林水産技術「研究ジャーナル」. 印刷中. (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 民谷栄一: "機器分析ガイドブック" 丸善(印刷中), (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] E.Tamiya: "Olfaction and Taste XI" Springer-Verlag, (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Murakami,Y.: "Integration of Enzyme-Immobilized Column with Electrochemical Flow Cell,Using Micromachining Techniques for a Glucose Defection System" Anal.Chem.65. 2731-2735 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Matsui,J.: "Molecular Recognition in Continuous Polymer Rods Prepared by a Molecular Imprinting Technique" Anal.Chem.65. 2223-2224 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Song,M.I.: "Alternating current field enhanced latex immunoassay for human myglobin as measured by image avolysis" Anal.Chim.Acta. 282. 193-198 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Chen.C.Y.: "Biocompatble Needle-Type Glucose Sensor with Potential for Use In Vivo" Electroanalysis. 5. 269-276 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Navera,E.N.: "Micro-choline sensor for acetylcholineesterase determination" Anal.Chim.Acta. 281. 673-679 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tamiya,E.: "Ultra micro glutamate sensor using platinized carbon-fiber electrode and integrated countei electrode" Sensors & Actuafors. B10. 179-184 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hirose,A.: "Optics in Medicine,Biology and Environmental Research" Elsevier Science Publishers, 285 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tamiya,E.: "Bioproducts and Bioprocesses" Springer-Verlag, 309 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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