Dual mature microRNA-responsive logic biosensing platform based on CRISPR/Cas12a and DNA nanocage

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Cancer Research

Background:

• Mature microRNAs are vital in cancer development but face challenges as biomarkers due to precursor interference and false positives.
• Existing methods struggle with specificity and accuracy in detecting microRNAs for cancer diagnosis.

Purpose of the Study:

• To develop a dual mature microRNA-responsive second-order (YES-AND) logic biosensing platform for precise cancer diagnosis.
• To overcome limitations of current microRNA biomarker detection, enhancing accuracy and reducing false signals.

Main Methods:

• Utilized DNA nanocages as "YES" gates for size-selective microRNA discrimination via strand displacement.
• Employed CRISPR/Cas12a system as an "AND" gate, requiring dual activators for signal amplification.
• Applied the platform to detect non-small cell lung cancer-related microRNAs in clinical serum samples.

Main Results:

• Achieved high sensitivity with a limit of detection (LOD) of 100 pM.
• Demonstrated excellent trueness with a recovery rate of ≥90%.
• Successfully differentiated between mature microRNAs and precursor microRNAs, improving diagnostic specificity.

Conclusions:

• The developed second-order logic biosensing platform offers accurate and reliable cancer diagnosis.
• This intelligent biosensor advances complex computations within pathological networks for biological and clinical applications.
• Represents a significant step towards developing sophisticated biosensors for disease diagnosis.