Analytical chemistrySep 20, 2025

Highly Sensitive Detection of Lung Cancer Biomarker miRNA-21 in Tiny Particles Using Enhanced CRISPR and SERS Technology

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jan 3, 2026

Abstract

The assay achieved a limit of detection as low as 0.62 aM for exosomal miRNA-21.

Exosomal miRNA-21 is associated with tumor progression in early-stage lung cancer.
Challenges in detecting miRNA-21 include its low abundance, short sequence length, and high-sequence similarity.
The developed platform combines rolling circle amplification with CRISPR technology for enhanced detection.
The assay successfully discriminated miRNA-21 from various single- and multinucleotide variants.
In a preliminary cohort of 20 early-stage lung cancer patients and 20 healthy controls, the method achieved 100% sensitivity and specificity.

Simplified

Exosomal miRNA-21 has emerged as a promising biomarker for early-stage lung cancer due to its close association with tumor progression and its stability in circulation. However, its low abundance, short sequence length, and high-sequence similarity present significant detection challenges. To address this, we developed an ultrasensitive surface-enhanced Raman scattering (SERS) platform that integrates rolling circle amplification (RCA) with clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 12a (Cas12a) for the detection of exosomal miRNA-21. RCA provides target-dependent amplification with stringent sequence discrimination via padlock probe ligation, while the CRISPR/Cas12a system facilitates robust signal generation through trans-cleavage activity. The final SERS readout enables molecular-level sensitivity by detecting nanotag-labeled cleavage events. The assay achieved a limit of detection as low as 0.62 aM and effectively discriminated miRNA-21 from multiple single- and multinucleotide variants. As a proof of concept, we applied this method to the detection of exosomal miRNA-21 extracted from the serum of 20 early-stage lung cancer patients and 20 healthy controls, achieving 100% sensitivity and 100% specificity (AUC = 1.0) in this preliminary cohort. These findings demonstrate the strong potential of the RCA-CRISPR-SERS platform for noninvasive early-stage lung cancer diagnosis based on exosomal miRNA-21 detection.

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Highly Sensitive Detection of Lung Cancer Biomarker miRNA-21 in Tiny Particles Using Enhanced CRISPR and SERS Technology

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