Redesigning CRISPR-Cas12a into a versatile biosensor with customizable RNA guides

The engineered CRISPR-Cas12a system achieves a detection limit of 0.5 aM for synthetic circHER2 RNA.

• The platform allows direct RNA detection without the need for reverse transcription.
• It features an autocatalytic circuit that enhances signal amplification.
• The system demonstrates high specificity, capable of discriminating single-nucleotide variants in DNA with a selectivity of up to 908.7.
• Robust performance is validated by accurately assessing circHER2 levels in breast cancer cell lines within complex cellular environments.
• This approach re-engineers Cas12a to overcome its limitations and establishes a new framework for molecular diagnostics.

Simplified