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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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Development and application of G4-Flame as a visual biosensor for G4-DNA.

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  • 1Department of Hepatobiliary Surgery, State Key Laboratory of Immune Response and Immunotherapy, Centre for Leading Medicine and Advanced Technologies of IHM, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230001, China.

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Summary

Scientists developed G4-Flame, a biosensor to visualize G-quadruplex DNA (G4-DNA) in real-time. Elevated G4-DNA levels in cancer patients suggest its potential as an early detection biomarker.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biochemistry

Background:

  • G-quadruplex DNA (G4-DNA) is a crucial noncanonical DNA structure involved in genomic regulation and disease.
  • Existing G4-DNA detection methods lack specificity, spatiotemporal resolution, and live-cell compatibility.

Purpose of the Study:

  • To develop a novel biosensor for real-time, high-resolution visualization of G4-DNA dynamics in living systems.
  • To investigate the spatiotemporal distribution and function of G4-DNA during the cell cycle and in disease states.

Main Methods:

  • Engineered G4-Flame, a genetically encoded fluorescent biosensor using circularly permuted YFP (cpYFP) and a G4-specific binding domain.
  • Utilized G4-Flame to visualize G4-DNA dynamics in living cells and analyzed serum samples.

Main Results:

  • G4-Flame demonstrated real-time, high-resolution visualization of G4-DNA with specificity across conformations.
  • Observed distinct nuclear and mitochondrial G4-DNA patterns during the cell cycle, with nuclear G4-DNA peaking in S phase.
  • Mitochondrial G4-DNA was found to suppress mitochondrial-encoded gene expression.
  • Significantly elevated serum G4-DNA levels were detected in cancer patients compared to healthy controls.

Conclusions:

  • G4-Flame is a transformative tool for studying G4-DNA spatiotemporal regulation in live systems.
  • G4-DNA holds potential as a biomarker for early cancer detection, bridging basic research and clinical applications.