Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

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...
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...
Experimental RNAi02:15

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Ectonucleotidases CD39 and CD73 expression levels are independent and inverse predictors of survival in muscle-invasive bladder cancer.

The journal of pathology. Clinical research·2026
Same author

Federated Propensity Score Matching for Bias Correction in ICU.

Studies in health technology and informatics·2026
Same author

What Is My Data Capable of? Using Performance Limits to Assess Data Quality.

Studies in health technology and informatics·2026
Same author

Assessing AI-Based Decision Support in Early Sepsis and AKI Recognition.

Studies in health technology and informatics·2026
Same author

Predicting blood transfusion after ICU admission in five databases: A comparison of three machine learning paradigms.

Digital health·2026
Same author

Comparison of a 3D-printed skin model with established methods for teaching punch biopsy with suturing.

Journal der Deutschen Dermatologischen Gesellschaft = Journal of the German Society of Dermatology : JDDG·2026
Same journal

Mapping the 3D Chromosome Organization of a Biosynthetic Gene Cluster by Capture Hi-C (CHi-C).

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Mapping the 3D Chromosome Organization of Streptomyces by Hi-C.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

CUT&Tag Epigenomic Profiling of Biosynthetic Gene Clusters in Arabidopsis thaliana.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Rhizobium rhizogenes-Mediated Hairy Root Transformation Protocol for Lotus japonicus and Other Legumes.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Characterization of Bioactive Saponins from Sea Cucumbers.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Methods for Functional Validation of Terpenoid Metabolic Clusters in Nicotiana benthamiana and Aspergillus oryzae.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

Related Experiment Video

Updated: May 18, 2026

mirMachine: A One-Stop Shop for Plant miRNA Annotation
06:16

mirMachine: A One-Stop Shop for Plant miRNA Annotation

Published on: May 1, 2021

Experimental miRNA target validation.

Jens Heyn1, Ludwig Christian Hinske, Carola Ledderose

  • 1Clinic of Anesthesiology, Clinic of the University of Munich, Munich, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|September 26, 2012
PubMed
Summary
This summary is machine-generated.

This study details a method for validating microRNA (miRNA) target interactions. The protocol involves creating a dual-luciferase vector and using a luciferase assay to confirm gene regulation by miRNAs.

More Related Videos

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

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
09:06

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

Published on: October 7, 2025

Related Experiment Videos

Last Updated: May 18, 2026

mirMachine: A One-Stop Shop for Plant miRNA Annotation
06:16

mirMachine: A One-Stop Shop for Plant miRNA Annotation

Published on: May 1, 2021

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

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
09:06

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

Published on: October 7, 2025

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) are key regulators of gene expression.
  • Investigating cellular regulatory circuits often involves studying miRNAs.
  • Computational methods predict miRNA-target interactions, necessitating experimental validation.

Purpose of the Study:

  • To present a reliable protocol for validating microRNA (miRNA) target gene interactions.
  • To provide a method for confirming the functional activity of predicted miRNA-target relationships.

Main Methods:

  • Cloning a dual-luciferase miRNA target expression vector.
  • Transfecting cells with the constructed vector and a precursor miRNA (pre-miRNA).
  • Performing a luciferase assay to quantify miRNA-mediated gene regulation.

Main Results:

  • The described protocol enables the experimental validation of miRNA-target interactions.
  • Successful transfection and luciferase assays confirm the regulatory role of specific miRNAs on target genes.

Conclusions:

  • This chapter offers a practical and effective method for the biomolecular validation of miRNA-target interactions.
  • The presented protocol is crucial for advancing the understanding of miRNA-mediated gene regulation in cellular processes.