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Related Concept Videos

MicroRNAs01:22

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...
MicroRNAs01:22

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

Updated: May 25, 2026

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
11:00

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

Published on: June 12, 2018

A functional assay for microRNA target identification and validation.

Joop Gäken1, Azim M Mohamedali, Jie Jiang

  • 1Department of Haematological Medicine, King's College London, Rayne Institute, London SE5 9NU, UK. joop.gaken@kcl.ac.uk

Nucleic Acids Research
|February 11, 2012
PubMed
Summary

Identifying microRNA targets is challenging. This study introduces a novel functional assay using a 3'-UTR-enriched library and selection strategy to accurately identify biologically relevant microRNA targets.

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Last Updated: May 25, 2026

Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
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Published on: June 12, 2018

An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
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Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library
08:40

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

Published on: April 6, 2012

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) are small RNA molecules regulating gene expression by binding to target mRNAs.
  • Identifying specific miRNA targets is difficult due to incomplete sequence homology, often relying on predictive algorithms.
  • Accurate target identification is crucial for understanding miRNA function in cellular processes.

Purpose of the Study:

  • To develop and validate a novel functional assay for the identification of biologically relevant miRNA targets.
  • To overcome the limitations of computational prediction methods for miRNA target discovery.
  • To provide a robust method for confirming miRNA-target interactions.

Main Methods:

  • Development of a functional assay utilizing a 3 -UTR-enriched library.
  • Implementation of a positive/negative selection strategy for target enrichment.
  • Proof-of-principle validation using miR-130a and its known target MAFB.

Main Results:

  • Successful identification of MAFB and five additional novel targets for miR-130a.
  • Confirmation of identified targets through western blot analysis.
  • Validation of target interactions via knockdown experiments.

Conclusions:

  • The novel functional assay is effective for the accurate identification of miRNA targets.
  • This method overcomes challenges associated with computational miRNA target prediction.
  • The validated strategy enables robust functional identification of miRNA targets in biological systems.