Homogeneous assay of target molecules based on chemiluminescence resonance energy transfer (CRET) using DNAzyme-linked aptamers

Hyoyoung Mun, Eun Jung Jo, Taihua Li, Hyou Arm Joung, Dong Gu Hong, Won Bo Shim, Cheulhee Jung, Min Gon Kim

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

We have designed a single-stranded DNAzyme-aptamer sensor for homogeneous target molecular detection based on chemiluminescence resonance energy transfer (CRET). The structure of the engineered single-stranded DNA (ssDNA) includes the horseradish peroxidase (HRP)-like DNAzyme, optimum-length linker (10-mer-length DNA), and target-specific aptamer sequences. A quencher dye was modified at the 3' end of the aptamer sequence. The incorporation of hemin into the G-quadruplex structure of DNAzyme yields an active HRP-like activity that catalyzes luminol to generate a chemiluminescence (CL) signal. In the presence of target molecules, such as ochratoxin A (OTA), adenosine triphosphate (ATP), or thrombin, the aptamer sequence was folded due to the formation of the aptamer/analyte complex, which induced the quencher dye close to the DNAzyme structure. Consequently, the CRET occurred between a DNAzyme-catalyzed chemiluminescence reaction and the quencher dye. Our results showed that CRET-based DNAzyme-aptamer biosensing enabled specific OTA analysis with a limit of detection of 0.27. ng/mL. The CRET platform needs no external light source and avoids autofluorescence and photobleaching, and target molecules can be detected specifically and sensitively in a homogeneous manner.

Original languageEnglish
Pages (from-to)308-313
Number of pages6
JournalBiosensors and Bioelectronics
Volume58
DOIs
Publication statusPublished - 2014 Aug 15
Externally publishedYes

Keywords

  • Aptamer
  • Aptasensor
  • Chemiluminescence resonance energy transfer
  • DNAzyme
  • Quenching

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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