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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Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

PubMed

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

Abstract

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Abstract

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