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Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
Published on: October 6, 2022
Aptamer selection based on G4-forming promoter region
Wataru Yoshida1, Taiki Saito, Tomomi Yokoyama
1Department of Biotechnology and Life Science, Tokyo University of Agriculture & Technology, Koganei, Tokyo, Japan.
Plos One
|June 11, 2013
Summary
We developed G4 promoter-derived aptamer selection (G4PAS), a novel method for identifying DNA aptamers. This technique utilizes G-quadruplex (G4) DNA structures found in gene promoter regions to discover aptamers targeting specific proteins like VEGF165, PDGF-AA, and RB1.
Area of Science:
- Biotechnology
- Molecular Biology
- Genomics
Background:
- Aptamers are nucleic acid ligands with high specificity for target molecules.
- G-quadruplex (G4) structures are potential scaffolds for aptamer development.
- Gene promoter regions often contain G4-forming sequences.
Purpose of the Study:
- To develop a novel method for aptamer identification without in vitro selection.
- To investigate the potential of G4 DNA structures in promoter regions as aptamers.
- To establish G4 promoter-derived aptamer selection (G4PAS) for aptamer discovery.
Main Methods:
- Developed the G4 promoter-derived aptamer selection (G4PAS) method.
- Applied G4PAS to identify aptamers against VEGF165, PDGF-AA, and RB1.
- Utilized Surface Plasmon Resonance (SPR) for binding analysis and dissociation constant (Kd) determination.
Main Results:
- Successfully identified DNA aptamers for VEGF165, PDGF-AA, and RB1 using G4PAS.
- Determined Kd values for VEGF165, PDGF-AA, and RB1 aptamers: 1.7 × 10⁻⁷ M, 6.3 × 10⁻⁹ M, and 4.4 × 10⁻⁷ M, respectively.
- Demonstrated G4PAS as a simple and rapid aptamer identification technique.
Conclusions:
- G4PAS is an effective and efficient method for aptamer discovery.
- G4 DNA structures in promoter regions can serve as aptamers against their gene products.
- G4PAS has broad applicability for identifying aptamers against various target proteins with promoter G4 DNA sequences.
