In-solution multiplex miRNA detection using DNA-templated silver nanocluster probes

Area of Science:

• Biochemistry
• Nanotechnology
• Molecular Biology

Background:

• MicroRNAs (miRNAs) are crucial regulatory molecules and potential biomarkers for disease diagnosis.
• Existing miRNA detection methods often lack speed, simplicity, or multiplexing capabilities.
• Developing rapid, sequence-selective detection methods for miRNAs is a significant challenge.

Purpose of the Study:

• To advance a DNA/silver nanocluster (AgNCs)-based fluorescence method for multiplex microRNA detection in solution.
• To design and investigate spectrally distinct fluorescent DNA/AgNCs probes for simultaneous miRNA identification.
• To demonstrate a proof of concept for a rapid, one-step, in-solution multiplex miRNA detection system.

Main Methods:

• Tethering of DNA-12nt scaffolds with 9 different AgNCs emitters to target-sensing DNA sequences.
• Investigating the role of DNA scaffolds and target-sensing sequences in tuning fluorescence emission.
• Utilizing DNA/AgNCs probes to achieve green, red, and near-infrared (NIR) fluorescence signals.

Main Results:

• Successfully created spectrally distinct DNA/AgNCs probes by varying both AgNC emitters and tethered DNA sequences.
• Achieved three distinct fluorescence emissions (green, red, NIR) from the developed probes.
• Demonstrated the capability of these probes for multiplex miRNA detection in a single reaction.

Conclusions:

• The developed DNA/AgNCs probes enable tunable fluorescence emission across the visible to NIR spectrum.
• This approach offers a rapid, one-step, and sequence-selective method for multiplex miRNA detection in solution.
• The study presents a promising platform for sensitive and efficient biomarker analysis.