Fluorescence detection of DNA, adenosine-5'-triphosphate (ATP), and telomerase activity by zinc(II)-protoporphyrin IX/G-quadruplex labels

Anal Chem. 2012 Jun 5;84(11):4789-97. doi: 10.1021/ac300348v. Epub 2012 May 22.

Abstract

The zinc(II)-protoporphyrin IX (ZnPPIX) fluorophore binds to G-quadruplexes, and this results in the enhanced fluorescence of the fluorophore. This property enabled the development of DNA sensors, aptasensors, and a sensor following telomerase activity. The DNA sensor is based on the design of a hairpin structure that includes a "caged" inactive G-quadruplex sequence. Upon opening the hairpin by the analyte DNA, the resulting fluorescence of the ZnPPIX/G-quadruplex provides the readout signal for the sensing event (detection limit 5 nM). Addition of Exonuclease III to the system allows the recycling of the analyte and its amplified analysis (detection limit, 200 pM). The association of the ZnPPIX to G-quadruplex aptamer-substrate complexes allowed the detection of adenosine-5'-triphosphate (ATP, detection limit 10 μM). Finally, the association of ZnPPIX to the G-quadruplex repeat units of telomers allowed the detection of telomerase activity originating from 380 ± 20 cancer 293T cell extract.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / analysis*
  • Biosensing Techniques*
  • DNA / analysis*
  • Exodeoxyribonucleases / chemistry
  • Exodeoxyribonucleases / metabolism
  • Fluorescence
  • Fluorescent Dyes
  • G-Quadruplexes
  • HEK293 Cells
  • Humans
  • Limit of Detection
  • Protoporphyrins / chemistry*
  • Telomerase / analysis*
  • Telomerase / metabolism

Substances

  • Fluorescent Dyes
  • Protoporphyrins
  • zinc protoporphyrin
  • Adenosine Triphosphate
  • DNA
  • Telomerase
  • Exodeoxyribonucleases
  • exodeoxyribonuclease III