Orderly nucleic acid aggregates by electrostatic self-assembly in single cells for miRNA detection and visualizing

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Biology

Background:

• Mesoporous silica nanoparticles (MSNs) are versatile platforms for biomolecule conjugation.
• Developing non-destructive methods for intracellular analysis is crucial for understanding cellular processes.
• MicroRNA (miRNA) detection is vital for disease diagnosis and biological research.

Purpose of the Study:

• To develop a novel method for in situ detection and imaging of miRNAs within living cells.
• To create orderly nucleic acid aggregates (ONAAs) self-assembled on functionalized nanoparticles.
• To utilize electrostatic DNA self-assembly and hybridization chain reaction (HCR) for signal amplification.

Main Methods:

• Synthesis of mesoporous silica nanoparticles (MSNs) functionalized with positively charged aminopropyl groups (PC).
• Self-assembly of orderly nucleic acid aggregates (ONAAs) on the surface of PCMSNs.
• Implementation of a hybridization chain reaction (HCR) on the PCMSN surface for signal amplification.
• In situ detection and fluorescence imaging of miRNAs in living cells.

Main Results:

• Successfully developed ONAAs self-assembled on PCMSNs.
• Demonstrated a novel electrostatic DNA self-assembly for HCR on PCMSNs within single cells.
• Established a non-destructive amplification strategy for miRNA detection and in situ imaging.
• Achieved prominent and aggregated fluorescence-bright spots for sensitive miRNA visualization.

Conclusions:

• ONAAs-PCMSNs provide a powerful platform for non-destructive intracellular miRNA detection.
• The developed electrostatic DNA self-assembly and HCR strategy enables sensitive and specific miRNA imaging in living cells.
• This approach holds potential for advancing diagnostics and fundamental biological research.