Multiple microRNAs analysis based on DNAzyme cascade DNA fiber barcodes for cell typing

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Nanotechnology

Background:

• Accurate microRNA (miRNA) profiling is crucial for precision medicine but faces challenges like cross-reactivity, cost, and stability with existing methods.
• Current diagnostic tools often lack the sensitivity and specificity required for reliable tumor subtyping and cell classification.

Purpose of the Study:

• To develop a novel DNAzyme-powered DNA fiber barcode platform for sensitive, specific, and multiplexed miRNA detection.
• To overcome the limitations of existing miRNA detection methods for improved clinical diagnostics.

Main Methods:

• Utilized a DNAzyme-based system combining G-quadruplex (G4)/hemin catalysis with miRNA-responsive displacement.
• Incorporated polydopamine (PDA)-mediated fluorescence modulation for signal generation and quenching via Förster resonance energy transfer (FRET).
• Developed a target-dependent "switch" mechanism where miRNA presence preserves fluorescence.

Main Results:

• Achieved sensitive miRNA detection with low limits (5.50 pM) and a broad linear range (50 pM-100 nM).
• Demonstrated dual qualitative and quantitative analysis capabilities.
• Enabled one-pot multiplex miRNA detection using multicolor barcoding.
• Showed high concordance with quantitative polymerase chain reaction (qPCR) validation.

Conclusions:

• The DNAzyme-powered DNA fiber barcode platform offers a stable, tunable, and efficient solution for miRNA detection.
• This technology overcomes existing barriers, enabling robust miRNA-based cell typing and precision diagnostics.
• The platform holds significant potential for advancing molecular diagnostics and personalized medicine.