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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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Detection of Bacteria Using Fluorogenic DNAzymes
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Fluorogenic target competitors for developing label-free and sensitive folding-unswitching aptamer sensors.

Xingli Zeng1, Xiufang Tong1, Jiahui Chen1

  • 1Key Laboratory of the Ministry of Education for Advanced Catalysis Materials, College of Chemistry and Materials Science, Zhejiang Normal University, Jinhua, 321004, Zhejiang, China.

Analytica Chimica Acta
|October 13, 2024
PubMed
Summary

This study introduces label-free folding-unswitching aptamer sensors (FUAS) that offer higher sensitivity than traditional methods. These FUAS utilize fluorogenic target competitors, overcoming limitations of existing aptamer sensors for broader applications.

Keywords:
BerberineFluorogenic target competitorsFolding-unswitching aptamer sensorsIsoquinoline alkaloidsSensitivity

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Aptamers are crucial in diagnostics and therapeutics, with structure-switching aptamer sensors (SSAS) widely used for target detection.
  • Traditional SSAS require costly modifications and can suffer from reduced sensitivity due to fluorophore/quencher labeling and complex designs.

Purpose of the Study:

  • To develop a novel, label-free aptamer sensor platform with enhanced sensitivity.
  • To overcome the cost and sensitivity limitations associated with conventional structure-switching aptamer sensors.

Main Methods:

  • Developed label-free folding-unswitching aptamer sensors (FUAS) using fluorogenic target competitors.
  • Screened natural isoquinoline alkaloids, identifying berberine (BER) as an effective competitor for adenine-based targets.
  • Utilized BER's fluorescence, which is modulated by target competition for binding sites on the aptamer.

Main Results:

  • The developed FUAS demonstrated significantly higher sensitivity compared to existing SSAS.
  • Successfully applied FUAS for detecting ATP and adenosine deaminase in serum samples.
  • Validated FUAS for screening inhibitors of adenosine deaminase, proving platform reliability.

Conclusions:

  • Fluorogenic target competitors enable the development of label-free FUAS with superior sensitivity.
  • This approach offers a cost-effective and high-performance alternative to traditional SSAS.
  • The FUAS platform holds promise for diverse applications, including targeted drug delivery.