A reduction-triggered fluorescence probe for sensing nucleic acids

Bioconjug Chem. 2008 Jun;19(6):1219-26. doi: 10.1021/bc800014d. Epub 2008 May 14.

Abstract

We have developed a reduction-triggered fluorescence probe with a new fluorogenic compound derivatized from Rhodamine for sensing oligonucleotides. The chemistry to activate the compound involves the reaction between the azide group of rhodamine derivatives and the reducing reagents, with the fluorescence signal appearing after reduction of the azide group. The signal/background ratio of this fluorogenic compound reached 2100-fold enhancement in fluorescence intensity. Dithio-1,4-threitol or triphenylphosphine as reducing reagents were successfully utilized for this chemistry to be introduced into the DNA probe. The genetic detection requires that two strands of DNA bind onto target oligonucleotides, one probe carrying a reducible fluorogenic compound while the other carries the reducing reagents. The reaction proceeds automatically without any enzymes or reagents under biological conditions to produce a fluorescence signal within 10-20 min in the presence of target DNA or RNA. In addition, the probe was very stable under biological conditions, even such extreme conditions as pH 5 solution, pH 10 solution, or high temperature (90 degrees C) with no undesirable background signal. The probes were successfully applied to the detection of oligonucleotides at the single nucleotide level in solution and endogenous RNA in bacterial cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Azides / chemistry*
  • Escherichia coli K12 / cytology
  • Escherichia coli K12 / metabolism
  • Fluorescence
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / chemical synthesis
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism*
  • Humans
  • Indicators and Reagents / chemistry
  • Nucleic Acids / analysis*
  • Nucleic Acids / metabolism
  • Oligonucleotides / metabolism
  • Oxidation-Reduction
  • RNA, Ribosomal, 23S / metabolism
  • Rhodamines / chemistry*

Substances

  • Azides
  • Fluorescent Dyes
  • Indicators and Reagents
  • Nucleic Acids
  • Oligonucleotides
  • RNA, Ribosomal, 23S
  • Rhodamines