Assembly of DNA Probes into Superstructures for Dramatically Enhancing Enzymatic Stability and Signal-to-Background Ratio

ACS Sens. 2018 Dec 28;3(12):2702-2708. doi: 10.1021/acssensors.8b01247. Epub 2018 Dec 5.

Abstract

DNA fluorescent probes are versatile tools that are widely used for biological detection in tubes. Using DNA probes in living systems, however, represents a significant challenge because of the endogenous nuclease-induced DNA degradation and strong background fluorescence in complex biological environments. Here, we show that assembling DNA probes into core-satellite gold nanoparticle (AuNP) superstructures could unprecedentedly enhance enzymatic stability and reduce background interference. The embedded DNA probes are protected from interaction with nuclease, eliminating the enzymatic degradation. In the meantime, the AuNP superstructures show extremely high quenching efficiency (>98%) toward the embedded DNA probes, whose fluorescence can be instantly turned on by the target binding, resulting in high signal-to-background ratio. To demonstrate these distinct properties, we made use of the assembled nanoprobes to monitor the ATP levels under different stimuli in living cells. The assembly strategy leads to a new opportunity for accurately sensing targets in living systems.

Keywords: DNA probes; assembly; biological detection; enhanced enzyme resistance; gold nanoparticles.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / analysis
  • Animals
  • Aptamers, Nucleotide / chemistry
  • Aptamers, Nucleotide / toxicity
  • Carbocyanines / chemistry
  • Carbocyanines / toxicity
  • Cell Line, Tumor
  • DNA / chemistry*
  • DNA / toxicity
  • DNA Probes / chemistry*
  • DNA Probes / toxicity
  • Deoxyribonuclease I / chemistry
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / toxicity
  • Gold / chemistry
  • Gold / toxicity
  • Humans
  • Metal Nanoparticles / chemistry*
  • Metal Nanoparticles / toxicity
  • Mice
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / methods
  • Proof of Concept Study

Substances

  • Aptamers, Nucleotide
  • Carbocyanines
  • DNA Probes
  • Fluorescent Dyes
  • cyanine dye 5
  • Gold
  • Adenosine Triphosphate
  • DNA
  • Deoxyribonuclease I