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1988 Fiscal Year Final Research Report Summary

Development of site-specific RNA cleaving method using modified DNA splints-RNase H system

Research Project

Project/Area Number 62870091
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field Biological pharmacy
Research InstitutionHokkaido University

Principal Investigator

OHTSUKA Eiko  Faculty of Pharmaceutical Sciences, Hokkaido University, 薬学部, 教授 (80028836)

Co-Investigator(Kenkyū-buntansha) IWAI Shigenori  Faculty of Pharmaceutical Sciences, Hokkaido University, 薬学部, 助手 (10168544)
INOUE Hideo  Faculty of Pharmaceutical Sciences, Hokkaido University, 薬学部, 助手 (80088856)
Project Period (FY) 1987 – 1988
KeywordsSite-specific cleavage of RNA / Sequence-specific cleavage of RNA / RNase H / RNA・modified DNA hybrid / 2'-O-Methyloligoribonucleotide / Chimeric oligonucleotide / 大腸菌tRNA<met(\)f>
Research Abstract

The method for cleaving RNA site-specifically with RNase H was developed; RNase H is known to hydrolyze RNA only in RNA-DNA hybrids. The method invloves the use of the chimeric oligonucleotide containing a tetradeoxyribonucleotide and 2'-0-methyloligoribonucleotide(s) as a DNA splint.
Model experiments were carried out using the hybrids of oligo-ribonucleotides (9 and 18mer) and chimeric splints (9mer), and E. coli RNase H. We found that the ribo-strands were cleaved specifically at the 3'-end of the RNA DNA hybrid regions. This technique was successfully applied to the site-directed cleavage of the loop and stem regions in RNA 90mer. We also examined applicability of the method using E. coli tRNA (f Met) and found that the cleavages occurred at desired positions in the acceptor-stem region.

  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] Hideo Inoue: Nucleic Acids Research. 15. 6131-6148 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideo Inoue: FEBS Letters. 215. 327-330 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Susumu Shibahara: Nucleic Acids Research. 15. 4403-4415 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideo Inoue: Nucleic Acids Research Symposium Series. NO.19. 135-138 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Eiko Ohtsuka: Seikagaku. 60. 339-347 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideo,Inoue: "Synthesis and hybridization studies on two complementary nona(2'-O-methyl)ribonucleotides" Nucleic Acids Research. 15. 6131-6148 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hideo,Inoue: "Sequence-dependent hydrolysis of RNA using modified oligonucleotide splints and RNase H" FEBS Letters. 215. 327-330 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Susumu,Shibahara: "Site-directed cleavage of RNA" Nucleic Acids Research. 15. 4403-4415 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hideo,Inoue: "Sequence-specific cleavage of RNA using chimeric DNA splints and RNase H" Nucleic Acids Research Symposium Series. No. 19. 135-138 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Eiko,Ohtsuka: "Synthesis of modified oligonucleotides and applications for the study of nucleic acids" Seikagaku. 60. 339-347 (1988)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1990-12-19  

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