Analytical methods : advancing methods and applicationsJan 19, 2026

Using a Bst enzyme to boost Cas12a detection of microRNA

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jan 20, 2026

Abstract

The assay detects microRNA-21 with a sensitivity down to 9.25 × 10^(-10) M.

An ultrasensitive fluorescence platform was developed for detecting microRNA-21 by combining target recycling and signal amplification.
The assay utilizes Bst DNA polymerase to enable efficient recycling of the target, enhancing the generation of DNA duplex activators.
Cas12a-mediated signal amplification results in significantly increased fluorescence responses.
The platform demonstrates excellent sequence selectivity for miR-21.
Successful application of the method was achieved in monitoring miR-21 in both cell lysates and clinical serum samples.

Simplified

Quantification of trace microRNAs is crucial for early disease diagnosis but remains technically challenging. Herein, we developed an ultrasensitive fluorescence platform for microRNA-21 (miR-21) detection by integrating Bst DNA polymerase - assisted target recycling with CRISPR/Cas12a-mediated signal amplification. In this design, the target miRNA triggers toehold-mediated opening of a hairpin probe, followed by Bst-driven primer extension that enables efficient target recycling and the generation of abundant DNA duplex activators. Subsequently, these activators induce strong-cleavage activity of Cas12a, producing markedly enhanced fluorescence responses. Benefiting from the dual amplification of enzymatic recycling and Cas12a activation, the proposed assay exhibits high sensitivity toward miR-21 with a detection limit down to 9.25 × 10M. Furthermore, the platform exhibited excellent sequence selectivity and was successfully applied to monitor miR-21 in both cell lysates and clinical serum samples. Considering its convenient operation, strong analytical performance, and simple readout mode, this method holds great potential for trace biomarker analysis in clinical diagnostics. trans -12

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Using a Bst enzyme to boost Cas12a detection of microRNA

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