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Related Concept Videos

MicroRNAs01:22

MicroRNAs

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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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MicroRNAs01:22

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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...
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Related Experiment Video

Updated: May 4, 2026

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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A highly sensitive and selective homogenous assay for profiling microRNA expression.

Huimin Deng1, Wei Shen1, Yuqian Ren1

  • 1Department of Chemistry, National University of Singapore, Singapore 117543, Singapore.

Biosensors & Bioelectronics
|December 17, 2013
PubMed
Summary

This study presents a highly sensitive microRNA (miRNA) assay using DNAzyme-capped probes and a duplex-specific nuclease for amplified colorimetric detection. The method enables sensitive miRNA detection down to subfemtomolar levels in real-world samples.

Keywords:
3,3′,5,5′-TetramethylbenzidineColorimetryDNAzymeDuplex-specific nucleaseMicroRNA

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • MicroRNAs (miRNAs) are crucial regulators of gene expression.
  • Sensitive and selective detection of miRNAs is essential for diagnostics and research.
  • Existing miRNA assay methods often face limitations in sensitivity, selectivity, or complexity.

Purpose of the Study:

  • To develop a highly sensitive and selective homogeneous assay for microRNA detection.
  • To enable direct profiling of miRNAs in complex biological samples.
  • To achieve colorimetric detection of miRNAs at subfemtomolar levels.

Main Methods:

  • Utilized DNAzyme-capped capture probes (DZ-CPs) immobilized on magnetic beads.
  • Employed a duplex-specific nuclease (DSN) for cleavage of DNAzyme moieties upon miRNA hybridization.
  • Developed an amplified colorimetric detection strategy using the cleaved DNAzyme moieties and TMB substrate.

Main Results:

  • Achieved homogeneous detection of miRNAs with high sensitivity and selectivity.
  • Demonstrated colorimetric detection of miRNAs down to subfemtomolar concentrations.
  • The assay showed exceptional amplification power due to DSN activity and cumulative signal generation.
  • The isothermal amplification and simple protocol facilitated direct miRNA profiling in real-world samples.

Conclusions:

  • The developed assay offers a powerful tool for sensitive and selective miRNA quantification.
  • The method's simplicity and sensitivity make it suitable for direct analysis of miRNAs in various biological samples.
  • This assay has significant potential for applications in molecular diagnostics and biomedical research.