Biosensors & bioelectronicsJan 23, 2026

DNA-triggered CRISPR/Cas12a circuit for highly sensitive and flexible biosensing

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jan 24, 2026

Abstract

The CBD biosensing platform achieves a detection limit of 14 CFU/mL for methicillin-resistant Staphylococcus aureus.

A bulge DNA (BD)-driven activation circuit enhances the sensitivity of the CRISPR/Cas12a biosensing tool.
For nucleic acid targets, the system initiates a positive feedback loop that amplifies detection signals.
Detection of pesticides and mycotoxins is possible at the picogram-per-milliliter level through a universal single-stranded DNA activator.
An 'OR' logic gate allows for simultaneous detection of two mycotoxins, demonstrating multiplexing capabilities.
The CBD platform shows versatility for both nucleic acid and non-nucleic acid target detection.

Simplified

CRISPR/Cas12a has emerged as an innovative biosensing tool; however, its intrinsically linear accumulation of cleavage signals limits detection sensitivity. Herein, we report a bulge DNA (BD)-driven CRISPR/Cas12a dynamic activation circuit, termed CBD, as a highly sensitive and versatile biosensing platform for both nucleic acid and non-nucleic acid targets. The BD structure was rationally engineered to undergo programmable structural and functional transformation upon bulge degradation, thereby initiating exponential, self-amplifying activation of the Cas12a circuit. For nucleic acid targets, direct Cas12a recognition triggers BD cleavage and a positive feedback loop, enabling highly sensitive detection with a limit of 14 CFU/mL for methicillin-resistant Staphylococcus aureus. For non-nucleic acid targets, a universal single-stranded DNA activator was linked to the aptamer-complementary strand, enabling target-responsive release and subsequent initiation of the CBD system without altering the crRNA or BD sequence. This strategy enabled the detection of pesticides and mycotoxins at the picogram-per-milliliter level. Furthermore, an "OR" logic gate was constructed for the simultaneous detection of dual mycotoxins, highlighting the platform's capability for multiplexed hazard monitoring. Overall, CBD demonstrates significant potential as a new paradigm for next-generation biosensing technologies.

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DNA-triggered CRISPR/Cas12a circuit for highly sensitive and flexible biosensing

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