DNA Gate-Based CRISPR-Cas Exponential Amplification System for Ultrasensitive Small Extracellular Vesicle Detection to Enhance Breast Cancer Diagnosis

The DNA gate-based exponential amplification CRISPR-Cas system achieved a detection limit as low as 1.02 × 10 particles/mL for tumor-derived extracellular vesicles.

• Tumor-derived extracellular vesicles (tEVs) may serve as potential biomarkers for noninvasive cancer diagnosis due to their surface proteins related to parent cells.
• Phenotype heterogeneity and low abundance of tEVs present challenges for high-sensitivity multiplex detection.
• The developed DGEAC system integrates target recognition and cleavage activity for accurate detection of CD63 and vascular endothelial growth factor (VEGF) proteins on tEVs.
• The system can distinguish tEVs from different breast cancer cell lines, including MDA-MB-231, MCF-7, SKBR3, and MCF-10A.
• The DGEAC system demonstrated improved differentiation of breast cancer stages with an area under the curve (AUC) of 98.3%, compared to 90.0% for linear amplification.

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