Allosteric Genetically Encoded Dual-Color Fluorogenic RNA Biosensor Simultaneously Illuminates miRNA and Its...

Area of Science:

• Biomolecular Engineering
• Molecular Diagnostics
• RNA Biology

Background:

• MicroRNAs (miRNAs) and their precursors (pre-miRNAs) are key biomarkers in physiological and pathological processes.
• Understanding their relationship is crucial for disease mechanism insights and early diagnosis.
• Existing detection methods often require complex procedures or labels.

Purpose of the Study:

• To develop a simple, sensitive, and label-free biosensor for simultaneous detection of miRNA and pre-miRNA.
• To enable monitoring of these nucleic acids in complex biological samples like cells and tissues.
• To establish a versatile platform for nucleic acid detection.

Main Methods:

• Construction of a dual-color fluorogenic RNA biosensor utilizing a hairpin allosteric-driven transcription circuit.
• Design of a multifunctional hairpin probe for target recognition, transcription amplification, and signal transduction.
• Utilizing aptamer-fluorophore complexes for signal generation upon target binding.

Main Results:

• The biosensor achieved isothermal, one-step, homogeneous detection of miR-155 and pre-miR-155 with attomolar sensitivity.
• Accurate quantification and single-cell level analysis of miRNA and pre-miRNA were demonstrated.
• Differential expression profiles between breast cancer and healthy tissues were successfully discriminated.

Conclusions:

• The developed RNA biosensor offers a powerful tool for sensitive and specific detection of miRNAs and pre-miRNAs.
• This platform provides a new paradigm for precise nucleic acid diagnostics and disease research.
• The biosensor's scalability allows for adaptation to detect various other nucleic acid targets.