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MicroRNAs

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...

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Updated: Dec 16, 2025

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as A Novel Detection and Quantification Method

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as A Novel Detection and Quantification Method

Published on: October 7, 2025

A Cas12a-mediated cascade amplification method for microRNA detection.

Huan-Huan Sun¹, Fang He, Ting Wang

¹Laboratory of Biosystem and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. binchengyin@ecust.edu.cn bcye@ecust.edu.cn.

|July 2, 2020

Summary

This summary is machine-generated.

This study introduces a novel isothermal method for detecting microRNAs (miRNAs) using Cas12a enzyme activity. The technique offers high sensitivity and specificity for miRNA detection, crucial for cancer diagnostics.

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Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Published on: April 14, 2015

Area of Science:

Biochemistry
Molecular Biology
Biotechnology

Background:

MicroRNAs (miRNAs) are critical regulators of biological processes.
miRNAs serve as vital biomarkers in cancer diagnosis, prognosis, and treatment.
Accurate and sensitive miRNA detection methods are essential for clinical applications.

Purpose of the Study:

To develop a highly sensitive and specific isothermal method for microRNA detection.
To leverage Cas12a trans-cleavage activity for signal amplification in miRNA detection.
To validate the method's performance in biological samples.

Main Methods:

Enzyme-assisted cascade amplification utilizing Cas12a trans-cleavage activity.
Target miRNA-initiated ligation reaction to generate transcription templates.
Transcriptional amplification of RNA strands, followed by DNAzyme-mediated crRNA generation.
Cas12a-activated trans-cleavage of DNA reporters for fluorescence signal generation.

Main Results:

Achieved a femtomolar limit of detection for target miRNAs.
Demonstrated high specificity in distinguishing homologous miRNA sequences.
Successfully applied the method for miRNA detection in various cell lines.

Conclusions:

The developed isothermal method provides a sensitive and specific platform for miRNA detection.
This Cas12a-based approach holds promise for clinical applications in cancer diagnostics and monitoring.
The enzyme-assisted cascade amplification strategy offers a robust tool for biomarker discovery and analysis.