A rapid, sensitive, and selective bioluminescence resonance energy transfer (BRET)-based nucleic acid sensing system

Manoj Kumar, Daohong Zhang, David Broyles, Sapna K. Deo

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

Here we report the design of a bioluminescence resonance energy transfer (BRET)-based sensing system that could detect nucleic acid target in 5. min with high sensitivity and selectivity. The sensing system is based on adjacent binding of oligonucleotide probes labeled with Renilla luciferase (Rluc) and quantum dot (Qd) on the nucleic acid target. Here Rluc, a bioluminescent protein that generates light by a chemical reaction, is employed as an energy donor, and a quantum dot is used as an energy acceptor. Bioluminescence emission of Rluc overlaps with the Qd absorption whereas the emission of Qd is shifted from the emission of Rluc allowing for monitoring of BRET. In the presence of target, the labeled probes bind adjacently in a head-to-head fashion leading to BRET from Rluc to Qd upon addition of a substrate coelenterazine. The sensing system could detect target nucleic acid in buffer as well as in Escherichia coli cellular matrix in 5. min with a detection limit of 0.54. pmol. The ability to detect target nucleic acid rapidly in a cellular matrix with high sensitivity will prove highly beneficial in biomedical and environmental applications.

Original languageEnglish (US)
Pages (from-to)133-139
Number of pages7
JournalBiosensors and Bioelectronics
Volume30
Issue number1
DOIs
StatePublished - Dec 15 2011

Keywords

  • Bioluminescence resonance energy transfer
  • Energy transfer
  • Nucleic acid sensing
  • Quantum dots
  • Renilla luciferase

ASJC Scopus subject areas

  • Biophysics
  • Biomedical Engineering
  • Biotechnology
  • Electrochemistry

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