Aptamer-based sensing platform using three-way DNA junction-driven strand displacement and its application in DNA...

Area of Science:

• Molecular Biology
• Biotechnology
• Nanotechnology

Background:

• DNA aptasensors offer sensitive and specific molecular detection.
• Label-free detection methods simplify assay design and reduce costs.
• Three-way DNA junctions provide a versatile scaffold for molecular interactions.

Purpose of the Study:

• To develop a novel aptasensor based on a three-way DNA junction for label-free fluorescence detection.
• To demonstrate the platform's ability to detect adenosine triphosphate (ATP) and DNA strands.
• To construct a molecular logic circuit using the aptasensor.

Main Methods:

• Fabrication of a label-free fluorescent sensing platform utilizing a three-way DNA junction.
• Incorporation of aptamer sequences into the DNA junction for target recognition.
• Employing split G-quadruplex enhanced fluorescence of protoporphyrin IX (PPIX) for signal readout.
• Mechanism investigation using native polyacrylamide gel electrophoresis.
• Construction of a logic circuit with ATP and DNA strands as inputs.

Main Results:

• The proposed three-way DNA junction-driven strand displacement mode was successfully implemented.
• The aptasensor demonstrated sensitive detection of ATP and DNA strands.
• A label-free fluorescence readout was achieved using PPIX.
• A functional molecular logic circuit was constructed, demonstrating ATP's role in modulating the DNA logic gate.

Conclusions:

• The developed aptasensor platform is sensitive and label-free.
• The platform can be adapted for detecting various targets by altering the aptamer sequence.
• This molecular platform holds potential for diverse applications in sensing and molecular computing.