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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...

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 ends...

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 ends...

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Related Experiment Video

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Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs

Published on: June 12, 2018

A quick reality check for microRNA target prediction.

¹The Biomedical Research Centre, University of British Columbia, 2222 Health Sciences Mall, Vancouver, BC, V6T 1Z3, Canada. kast@brc.ubc.ca

Expert Review of Proteomics

|April 20, 2011

Summary

Researchers developed a quantitative method to validate microRNA (miRNA) targets in organisms. This approach efficiently verifies known and identifies novel miRNA targets, advancing the study of gene regulation.

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Last Updated: Jun 2, 2026

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Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library

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Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells

Published on: September 28, 2011

Area of Science:

Biochemistry
Molecular Biology
Genetics

Background:

MicroRNA (miRNA)-mediated repression of mRNA translation is crucial for regulating protein abundance.
Predicting miRNA targets is challenging due to imperfect seed sequence matching and limited overlap among existing software tools.

Discussion:

Jovanovic et al. utilized a targeted, quantitative approach with selected reaction monitoring (SRM) assays to validate predicted miRNA targets.
SRM assays were developed and tested for numerous predicted let-7 targets and control genes in Caenorhabditis elegans.
The study highlights the feasibility of rapid, high-throughput assay development for miRNA target validation.

Key Insights:

SRM assays offer a powerful tool for verifying known and discovering novel miRNA targets at the organismal level.
The biological system and material significantly influence the observed frequency, extent, and direction of miRNA-mediated changes.
This quantitative method facilitates a deeper understanding of miRNA function in complex biological systems.

Outlook:

Selected reaction monitoring assays represent a promising new methodology for studying miRNA function across various organisms.
Further application of this technique can refine our understanding of post-transcriptional gene regulation.
This approach has the potential to accelerate the identification of miRNA roles in development and disease.