Visible sensing of nucleic acid sequences using a genetically encodable unmodified mRNA probe

Nucleic Acids Symp Ser (Oxf). 2006:(50):283-4. doi: 10.1093/nass/nrl141.

Abstract

We previously reported a molecular beacon-mRNA (MB-mRNA) strategy for nucleic acid detection/sensing in a cell-free translation system using unmodified RNA as a probe. Here in this presentation, we report that a combination with RNase H activity, which induces an additional process of irreversible cleavage of MB-domain, achieves an improved sequence selectivity (one nucleotide selectivity) and an enhanced sensitivity. This improved system finally enabled visible sensing of target nucleic acid sequence at a single nucleotide resolution under isothermal conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Biosensing Techniques*
  • Cell-Free System
  • Colorimetry
  • Fluorescence Resonance Energy Transfer
  • Genes, Reporter
  • Luciferases / analysis
  • Luciferases / genetics
  • Luminescent Agents / analysis
  • Luminescent Measurements
  • Nucleic Acid Hybridization / methods*
  • Nucleic Acids / analysis*
  • Oligodeoxyribonucleotides / analysis
  • Protein Biosynthesis*
  • RNA Probes / chemistry*
  • Receptors, CCR5 / genetics
  • Ribonuclease H*
  • beta-Galactosidase / analysis
  • beta-Galactosidase / genetics

Substances

  • Luminescent Agents
  • Nucleic Acids
  • Oligodeoxyribonucleotides
  • RNA Probes
  • Receptors, CCR5
  • Luciferases
  • Ribonuclease H
  • beta-Galactosidase