T7 exo-mediated FRET-breaking combined with DSN-RNAse-TdT for the detection of microRNA with ultrahigh signal-amplification

Analyst. 2019 May 13;144(10):3216-3220. doi: 10.1039/c9an00303g.

Abstract

A DSN-RNAse-TdT-T7 exo probing system allows the detection of miRNA 21 with very high sensitivity (LOD = 2.57 fM) and selectivity-the result of (i) avoiding the false-positive signal from miRNA reacting with TdT polymerase and (ii) signal amplification occurring through a FRET-breaking mechanism involving T7 exo.

MeSH terms

  • Bacteriophage T7 / enzymology
  • DNA Nucleotidylexotransferase / chemistry*
  • DNA Probes / chemical synthesis
  • DNA Probes / genetics
  • DNA, Single-Stranded / chemical synthesis
  • DNA, Single-Stranded / genetics
  • Exodeoxyribonucleases / chemistry*
  • Fluorescence Resonance Energy Transfer / methods
  • Fluorescent Dyes / chemistry
  • Humans
  • Limit of Detection
  • MicroRNAs / blood*
  • MicroRNAs / genetics
  • Nucleic Acid Amplification Techniques
  • Nucleic Acid Hybridization
  • RNA, Messenger / genetics
  • Ribonucleases / chemistry*

Substances

  • DNA Probes
  • DNA, Single-Stranded
  • Fluorescent Dyes
  • MIRN21 microRNA, human
  • MicroRNAs
  • RNA, Messenger
  • DNA Nucleotidylexotransferase
  • Exodeoxyribonucleases
  • Ribonucleases
  • gene b exonuclease