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Issue 28, 2020
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Integrating CRISPR-Cas12a with a DNA circuit as a generic sensing platform for amplified detection of microRNA

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Abstract

CRISPR-based diagnostics (CRISPR-Dx) has shown great promise in molecular diagnostics, but its utility in the sensing of microRNA (miRNA) biomarkers is limited by sensitivity, cost and robustness. Here, we describe a CRISPR-Dx method for the sensitive and cost-effective detection of miRNAs by rationally integrating CRISPR-Cas12a with DNA circuits. In this work, a modular catalytic hairpin assembly (CHA) circuit is designed to convert and amplify each target into multiple programmable DNA duplexes, which serve as triggers to initiate the trans-cleavage activity of CRISPR-Cas12a for further signal amplification. Such rational integration provides a generic assay for the effectively amplified detection of miRNA biomarkers. By simply tuning the variable regions in the CHA modules, this assay achieves sub-femtomolar sensitivity for different miRNA biomarkers, which improves the detection limit of CRISPR-Dx in the analysis of miRNA by 3–4 orders of magnitude. With the usage of the proposed assay, the sensitive assessment of miR-21 levels in different cancer cell lines and clinical serum samples has been achieved, providing a generic method for the sensitive detection of miRNA biomarkers in molecular diagnosis.

Graphical abstract: Integrating CRISPR-Cas12a with a DNA circuit as a generic sensing platform for amplified detection of microRNA

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Supplementary files

Article information


Submitted
02 Jun 2020
Accepted
17 Jun 2020
First published
19 Jun 2020

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2020,11, 7362-7368
Article type
Edge Article

Integrating CRISPR-Cas12a with a DNA circuit as a generic sensing platform for amplified detection of microRNA

S. Peng, Z. Tan, S. Chen, C. Lei and Z. Nie, Chem. Sci., 2020, 11, 7362
DOI: 10.1039/D0SC03084H

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    [Original citation] - Published by The Royal Society of Chemistry.

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