Sensitive detection of heart disease-related microRNAs using a CRISPR system controlled by a split promoter and catalytic hairpin assembly

A limit of detection as low as 38.9 aM for miRNA-155 is achieved within 90 minutes.

• The biosensing platform integrates catalytic hairpin assembly with a split-T7 promoter-regulated CRISPR/Cas12a system.
• The split-T7 promoter architecture allows for target-responsive reconstitution, generating abundant crRNA transcripts without the need for pre-synthesized exogenous crRNA.
• Triple cascade amplification is achieved through target recycling, transcription-driven signal enrichment, and specific fluorescence readout.
• The method is capable of discriminating single-base mismatches and performs reliably with clinical specimens from coronary heart disease patients.
• This approach reduces assay cost and complexity while maintaining excellent amplification efficiency.

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