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Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
Published on: April 4, 2025
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Aza-Macrocyclic Triphenylamine Ligands for G-Quadruplex Recognition
Isabel Pont1,2, Jorge González-García1,2, Mario Inclán1
1Department of Inorganic Chemistry, Institute of Molecular Science, University of Valencia, Catedrático José Beltran 2, 46980, Paterna, Spain.
Chemistry (Weinheim an Der Bergstrasse, Germany)
|May 17, 2018
Summary
New triphenylamine ligands selectively detect G-quadruplex (G4) DNA. The bi- and tri-branched ligands exhibit a strong "light-up" fluorescence effect upon binding, enabling sensitive G4 DNA detection.
Area of Science:
- Supramolecular Chemistry
- Bioorganic Chemistry
- Analytical Chemistry
Background:
- G-quadruplex (G4) DNA structures are crucial in biological processes and disease.
- Developing selective G4 DNA probes is essential for diagnostics and research.
- Triphenylamine derivatives offer versatile scaffolds for molecular recognition and sensing.
Purpose of the Study:
- To synthesize and characterize novel triphenylamine-based ligands with varying numbers of aza-macrocycles.
- To investigate the binding affinity and selectivity of these ligands towards G4 DNA compared to duplex DNA.
- To explore the potential of these ligands as fluorescent probes for G4 DNA detection.
Main Methods:
- Synthesis of triphenylamine-based ligands (TPA1PY, TPA2PY, TPA3PY).
- Physicochemical characterization using pH-metric titrations, UV/Vis, and fluorescence spectroscopy.
- G4 DNA binding and selectivity assessed via FRET melting assays, fluorimetric titrations, and circular dichroism spectroscopy.
Main Results:
- Successful synthesis of triphenylamine ligands with one, two, or three pendant aza-macrocycles.
- Demonstrated high affinity and selectivity of bi- and tri-branched ligands for G4 DNA over duplex DNA.
- Observed an intense, redshifted fluorescence emission ('light-up' effect) upon ligand-G4 DNA interaction, attributed to intermolecular aggregation.
Conclusions:
- Triphenylamine-based ligands, particularly TPA2PY and TPA3PY, are effective binders and selective sensors for G4 DNA.
- The observed fluorescence 'light-up' phenomenon provides a sensitive mechanism for G4 DNA detection.
- These ligands hold promise as advanced fluorescent probes for G-G4 DNA recognition in biological contexts.
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