Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

RNA Splicing

RNA Splicing

Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...

You might also read

Related Articles

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

Sort by

Same author

NuMA1 controls myonuclear motility in striated skeletal muscle through AMPK activity and is impaired in Duchenne muscular dystrophy.

Cell death & disease·2026

Same author

Knockdown of Gene Expression Following Activation of the NMD Pathway Using Splice-Switching Oligonucleotides.

Methods in molecular biology (Clifton, N.J.)·2025

Same author

SH3KBP1 promotes skeletal myofiber formation and functionality through ER/SR architecture integrity.

EMBO reports·2025

Same author

<i>RBM20</i> Gene in Patients With Cardiomyopathy: Phenotypic Expression for Loss-of-Function Versus Hotspot Variants.

Circulation. Heart failure·2025

Same author

Mobile Element Insertion in the APOB Exon 3 Coding Sequence: A New Challenge in Hypobetalipoproteinemia Diagnosis.

Clinical genetics·2024

Same author

Circulating Autoantibodies Targeting TREK-1 in Patients With Short-Coupled Ventricular Fibrillation.

Circulation·2024

Search research articles

Related Experiment Video

Updated: Sep 14, 2025

Using the E1A Minigene Tool to Study mRNA Splicing Changes

Using the E1A Minigene Tool to Study mRNA Splicing Changes

Published on: April 22, 2021

Functional Evaluation of Splice Variants Using a Minigene Strategy.

Angélique Nizou¹, Lana Mahfoud¹, Alexandre Janin²

¹UR20218 - NeurIT, University of Limoges, Limoges, France.

Methods in Molecular Biology (Clifton, N.J.)

|July 23, 2025

Summary

Evaluating genetic variants affecting RNA splicing is difficult without RNA. This study proposes using patient DNA with hybrid minigenes to assess splicing impacts, even when RNA is unavailable.

Keywords:

Bacterial transformation Cell transfection Homologous recombination-mediated cloning Hybrid minigene RNA splicing

More Related Videos

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

Identification of Alternative Splicing and Polyadenylation in RNA-seq Data

Identification of Alternative Splicing and Polyadenylation in RNA-seq Data

Published on: June 24, 2021

Related Experiment Videos

Last Updated: Sep 14, 2025

Using the E1A Minigene Tool to Study mRNA Splicing Changes

Using the E1A Minigene Tool to Study mRNA Splicing Changes

Published on: April 22, 2021

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

Identification of Alternative Splicing and Polyadenylation in RNA-seq Data

Identification of Alternative Splicing and Polyadenylation in RNA-seq Data

Published on: June 24, 2021

Area of Science:

Genetics
Molecular Biology
Bioinformatics

Background:

Assessing genetic variants impacting RNA splicing poses challenges, especially when RNA samples are unavailable or specific tissues are inaccessible.
Traditional methods like RNA sequencing require readily available RNA, limiting variant evaluation in certain contexts.

Purpose of the Study:

To present an alternative strategy for evaluating the splicing effects of genetic variants using only patient DNA.
To demonstrate the utility of the hybrid minigene system for assessing variant-induced splicing alterations.

Main Methods:

Amplifying the DNA region of interest from patient samples.
Cloning the amplified region into a hybrid minigene reporter system.
Transfecting the minigene construct into eukaryotic cells for functional analysis.

Main Results:

The hybrid minigene system allows for the estimation of splicing alterations caused by specific DNA variants.
Comparison between mutated and wild-type regions in transfected cells provides insights into variant impact.

Conclusions:

This DNA-based strategy offers a viable alternative for studying RNA splicing alterations when RNA is not accessible.
The hybrid minigene approach facilitates the evaluation of genetic alterations' impact on RNA splicing using readily available patient DNA.