Micropillar enhanced FRET-CRISPR biosensor for nucleic acid detection

Area of Science:

• Biotechnology
• Molecular Biology
• Microfluidics

Background:

• CRISPR technology offers high sensitivity and specificity for pathogen detection.
• High-aspect-ratio microstructures can enhance biochemical applications, but their use in CRISPR detection is underexplored.

Purpose of the Study:

• To develop a CRISPR-based biosensor using high-aspect-ratio microstructures for HPV 16 DNA detection.
• To optimize FRET assay performance and address microfluidic challenges.

Main Methods:

• Developed a FRET-based biosensor combined with high-aspect-ratio microstructures and Cas12a-mediated trans-cleavage.
• Investigated micropillar density effects on molecular binding and detection sensitivity.
• Evaluated surface chemical treatments and developed a method to mitigate bubble generation in microfluidics.

Main Results:

• Micropillars with higher density significantly enhanced molecular binding and increased detection sensitivity by tenfold.
• Two surface chemical treatment methods were assessed for FRET assay improvement.
• An effective method was developed to prevent bubble formation in microfluidic devices.

Conclusions:

• This work presents a novel micropillar-based approach for CRISPR-enabled viral detection.
• The findings offer valuable insights into optimizing microfluidic biochemical reactions for enhanced diagnostic sensitivity.