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G-Quadruplex Conformational Switching for miR-155-3p Detection Using a Ligand-Based Fluorescence Approach.

Pedro Lourenço1, Carla Cruz1,2

  • 1RISE-Health, Department of Medical Sciences, Faculty of Health Sciences, University of Beira Interior, Av. Infante D. Henrique, 6200-506 Covilhã, Portugal.

Biomolecules
|March 28, 2025
PubMed
Summary
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A novel molecular beacon (MB-G4) detects microRNA-155-3p (miR-155-3p) by forming a G-quadruplex structure. This method offers sensitive and specific detection of miR-155-3p, crucial for cancer biomarker applications.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • MicroRNA-155-3p (miR-155-3p) is a significant biomarker in diseases like cancer.
  • Sensitive and specific detection methods for miR-155-3p are essential for clinical applications.

Purpose of the Study:

  • To develop a novel molecular beacon (MB-G4) for sensitive and specific detection of miR-155-3p.
  • To characterize the conformational changes and optimize detection conditions for MB-G4.

Main Methods:

  • Utilized a molecular beacon (MB-G4) that forms a G-quadruplex (G4) structure upon binding to miR-155-3p.
  • Employed circular dichroism (CD) spectroscopy and native polyacrylamide gel electrophoresis (PAGE) for structural analysis.
  • Optimized experimental conditions (KCl and NMM concentrations) and evaluated detection limit (LOD) and specificity.
Keywords:
G-quadruplexlung cancermiR-155-3pmolecular beacon

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Last Updated: Jun 15, 2026

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Main Results:

  • MB-G4 demonstrated a conformational switch from hairpin to G4 structure upon hybridization with miR-155-3p.
  • The G4 structure was recognized by N-methyl mesoporphyrin IX (NMM), yielding a concentration-dependent fluorescence signal.
  • Achieved a low detection limit (LOD) of 10.85 nM with high specificity against miR-155-5p.
  • Successfully detected miR-155-3p in total RNA from A549 lung cancer cells.

Conclusions:

  • MB-G4 provides a sensitive and specific platform for miR-155-3p detection.
  • The G4 formation mechanism enables a reliable fluorescence-based assay.
  • Validated potential for analyzing biological samples, including those from cancer cell lines.