Is your ribozyme design really correct? A proposal of simple single turnover competition assay to evaluate ribozymes

Terumichi Tanaka, Osamu Inui, Natsuki Dohi, Noriko Okada, Hidechika Okada, Yo Kikuchi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Today, many nucleic acid enzymes are used in gene therapy and gene regulations. However, no simple assay methods to evaluate enzymatic activities, with which we judge the enzyme design, have been reported. Here, we propose a new simple competition assay for nucleic acid enzymes of different types to evaluate the cleaving efficiency of a target RNA molecule, of which the recognition sites are different but overlapped. Two nucleic acid enzymes were added to one tube to make a competition of these two enzymes for one substrate. The assay was used on two ribozymes, hammerhead ribozyme and hairpin ribozyme, and a DNA-enzyme. We found that this assay method is capable of application to those enzymes, as a powerful tool for the selection and designing of RNA-cleaving enzymes.

Original languageEnglish
Pages (from-to)1636-1644
Number of pages9
JournalBioscience, Biotechnology and Biochemistry
Volume65
Issue number7
DOIs
Publication statusPublished - 2001 Jul
Externally publishedYes

Keywords

  • 512 antigen mRNA
  • Competition assay
  • DNA-enzyme
  • Hairpin ribozyme
  • Hammerhead ribozyme

ASJC Scopus subject areas

  • Food Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Chemistry (miscellaneous)
  • Applied Microbiology and Biotechnology
  • Bioengineering

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