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A Cas12a-mediated cascade amplification method for microRNA detection

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Abstract

MicroRNAs (miRNAs) play a vital role in various biological processes and act as important biomarkers for clinical cancer diagnosis, prognosis, and therapy. Here, we took advantage of Cas12a trans-cleavage activity to develop an enzyme-assisted cascade amplification method for isothermal miRNA detection. A target miRNA-initiated ligation reaction would allow for the production of transcription templates that triggered the transcriptional amplification of RNA strands. These RNA strands were cleaved by the 8-17E DNAzyme to generate crRNAs and recycled RNAs which have the same sequence as the target miRNA. The amplified abundant crRNAs bound to Cas12a and dsDNA activators to form the complex, which trans-cleaved the ssDNA reporters to generate a fluorescence signal for miRNA quantitative analysis. The proposed method exhibits a femtomolar limit of detection and a good specificity in distinguishing the homologous sequences of miRNAs. Its practical application ability was further tested in different cell lines.

Graphical abstract: A Cas12a-mediated cascade amplification method for microRNA detection

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Article information


Submitted
20 Feb 2020
Accepted
09 Jun 2020
First published
16 Jun 2020

Analyst, 2020, Advance Article
Article type
Paper

A Cas12a-mediated cascade amplification method for microRNA detection

H. Sun, F. He, T. Wang, B. Yin and B. Ye, Analyst, 2020, Advance Article , DOI: 10.1039/D0AN00370K

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