DNAzyme-driven SDA reaction regulates CRISPR/Cas12a for highly sensitive and selective analysis of underexpressed miRNA

The biosensor demonstrated a linear detection range from 0.1 pmol/L to 1 nmol/L for underexpressed miRNA.

• A novel fluorescence biosensor was developed utilizing DNAzyme-driven strand displacement amplification and CRISPR/Cas12a for sensitive detection of underexpressed miRNA.
• In the presence of prostate cancer-associated miR-222, the biosensor showed a low fluorescent state due to the cleavage of substrates by intact DNAzymes.
• In contrast, the absence of miR-222 allowed for substrate activation, leading to a strong fluorescent signal through SDA and CRISPR/Cas12a activity.
• The biosensor achieved a detection limit of 33.5 fmol/L, indicating high sensitivity for identifying underexpressed miRNAs.
• Excellent specificity and anti-interference capacity were observed, enabling successful detection of miR-222 in blood samples.

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