Site-specific cleavage of natural mRNA sequences by newly designed hairpin catalytic RNAs

Yo Kikuchi*, Noriko Sasaki


研究成果: Article査読

19 被引用数 (Scopus)


The negative strand of tobacco ringspot virus satellite RNA is a self-cleaving RNA. Its catalytic domain and substrate domain have been identified, and the catalytic domain has been named hairpin catalytic RNA. Here we report the construction of a plasmid containing a modified hairpin catalytic RNA sequence that can be transcribed in vitro. Because this plasmid has two specific restriction enzyme recognition sites at both ends of the substrate binding site in the catalytic RNA sequence, it is possible to construct new plasmids by substituting different sequences in the substrate binding site. Using this plasmid, synthetic DNA, and in vitro transcription, we obtained three ribozymes designed to cleave Escherichia coli prolipoprotein signal peptidase (lsp) mRNA at specific sites. All three ribozymes cleaved the lsp mRNA sequence in vitro at the specific sites, and two of them cleaved it efficiently. Kinetic analyses showed that one had a higher kcat/Km value than that of the well-known hammerhead ribozyme. Problems associated with attaining the goal of expressing these ribozymes in vivo also are discussed.

ジャーナルNucleic Acids Research
出版ステータスPublished - 1991 12月 25

ASJC Scopus subject areas

  • 遺伝学
  • 統計学、確率および不確実性
  • 応用数学
  • 健康、毒物学および変異誘発
  • 毒物学
  • 遺伝学(臨床)


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