Live-Cell Sensing of Telomerase Activity by Using Hybridization-Sensitive Fluorescent Oligonucleotide Probes

Chembiochem. 2020 Apr 1;21(7):1022-1027. doi: 10.1002/cbic.201900555. Epub 2019 Dec 16.

Abstract

Live-cell sensing of telomerase activity with simple and efficient strategies remains a challenging target. In this work, a strategy for telomerase sensing by using hybridization-sensitive fluorescent oligonucleotide probes is reported. In the presence of telomerase and dNTPs, the designed supporting strand was extended and generated the hairpin structure that catalyzed the next telomerase extending reaction. The special extension mechanism increased the local concentration of another supporting strand and telomerase, which resulted in enhanced telomerase activity. The hybridization-sensitive oligonucleotide probes bound to the hairpin catalyst and generated turn-on fluorescence. This method realized the sensing of telomerase activity in HeLa cell extract with a detection limit below 1.6×10-6 IU μL-1 . The real-time in situ observation of telomerase extension was achieved in living HeLa cells. This strategy has been applied to monitor the efficiency of telomerase-targeting anticancer drugs in situ.

Keywords: drug discovery; fluorescence; imaging agents; oligonucleotides; telomerase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Fluorescent Dyes / chemistry*
  • HeLa Cells
  • Humans
  • Microscopy, Confocal
  • Nucleic Acid Hybridization
  • Oligonucleotide Probes / chemistry
  • Oligonucleotide Probes / metabolism*
  • Telomerase / metabolism*

Substances

  • Fluorescent Dyes
  • Oligonucleotide Probes
  • Telomerase