TalantaOct 26, 2026

Light-activated DNA nanorobot for ultra-sensitive one-step detection of two microRNAs in triple-negative breast cancer

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Oct 27, 2026

Abstract

The assay demonstrates a limit of detection of 48.7 aM for dual microRNA detection in triple-negative breast cancer.

MicroRNAs are identified as important biomarkers for early diagnosis of triple-negative breast cancer.
The innovative biosensing method utilizes a cascade DNA walker and CRISPR/Cas12a nanorobot for sensitive detection.
The assay operates effectively over a linear range from 100 aM to 1 pM.
Reliability is confirmed through serum recovery experiments and tests with cell lines.
Clinical validation on 30 plasma samples shows high diagnostic accuracy with an area under the curve of 0.964.

Simplified

MicroRNAs (miRNAs) are critical biomarkers for the early diagnosis of triple-negative breast cancer (TNBC), but sensitive simultaneous detection of multiple miRNAs remains a clinical challenge. Herein, we report an innovative biosensing method based on a cascade DNA walker-driven photo-controlled CRISPR/Cas12a nanorobot for one-pot, rapid, and ultrasensitive dual miRNA detection. Target miRNAs trigger the cascade DNA walker, which integrates catalytic hairpin assembly and enzyme-mediated strand displacement, to generate abundant single-stranded DNA (ssDNA). These ssDNA then activate the photo-controlled CRISPR/Cas12a nanorobot under light illumination, enabling amplified fluorescence signaling for dual-target quantification. The assay exhibits a linear range of 100 aM to 1 pM with a limit of detection of 48.7 aM. Serum recovery experiments and cell line tests confirm its reliability in complex matrices. Clinical validation using 30 plasma samples (15 TNBC patients, 15 healthy controls) demonstrates high diagnostic accuracy (area under the curve, AUC = 0.964). This method features simple operation and strong clinical translational potential, offering a reliable tool for the early diagnosis of TNBC.

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Light-activated DNA nanorobot for ultra-sensitive one-step detection of two microRNAs in triple-negative breast cancer

Abstract

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