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

RNA Splicing01:32

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...
RNA Splicing01:32

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...
Alternative RNA Splicing02:18

Alternative RNA Splicing

Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
Alternative RNA Splicing02:18

Alternative RNA Splicing

Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
Pre-mRNA Processing: RNA Splicing01:32

Pre-mRNA Processing: 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...
Chromatin Structure Regulates pre-mRNA Processing02:41

Chromatin Structure Regulates pre-mRNA Processing

In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...

You might also read

Related Articles

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

Sort by
Same author

Clinical and functional evaluation of non-missense MYH9 variants in MYH9-related disease.

Clinical and experimental nephrologyยท2026
Same author

Response to Comment on Tanabe et al. Prediction of Sarcopenia Onset in Type 2 Diabetes Using Urinary Titin Levels: A Japanese Prospective Cohort Study. Diabetes Care 2025;48:1844-1849.

Diabetes careยท2025
Same author

Myostatin antisense administration prevents sepsis-induced muscle atrophy and weakness in male mice.

Physiological reportsยท2025
Same author

Prediction of Sarcopenia Onset in Type 2 Diabetes Using Urinary Titin Levels: A Japanese Prospective Cohort Study.

Diabetes careยท2025
Same author

Serum titin/creatinine ratio as a biomarker for discriminating disease severity in Duchenne and Becker muscular dystrophies.

Frontiers in neurologyยท2025
Same author

Fragmented QRS in Lateral Leads on Electrocardiography Is Associated with Cardiac Dysfunction and Left Ventricular Dilation in Duchenne Muscular Dystrophy.

Biomedicinesยท2025

Related Experiment Video

Updated: May 23, 2026

Using the E1A Minigene Tool to Study mRNA Splicing Changes
10:25

Using the E1A Minigene Tool to Study mRNA Splicing Changes

Published on: April 22, 2021

Optimizing RNA/ENA chimeric antisense oligonucleotides using in vitro splicing.

Yasuhiro Takeshima1, Mariko Yagi, Masafumi Matsuo

  • 1Department of Pediatrics, Kobe University Graduate School of Medicine, Kobe, Japan. takesima@med.kobe-u.ac.jp

Methods in Molecular Biology (Clifton, N.J.)
|March 29, 2012
PubMed
Summary
This summary is machine-generated.

Researchers developed an efficient in vitro method to identify optimal antisense oligonucleotides (AOs) for Duchenne muscular dystrophy (DMD) exon skipping therapy. This system enables precise AO selection for molecular treatments targeting DMD.

More Related Videos

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
10:06

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
08:53

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

Related Experiment Videos

Last Updated: May 23, 2026

Using the E1A Minigene Tool to Study mRNA Splicing Changes
10:25

Using the E1A Minigene Tool to Study mRNA Splicing Changes

Published on: April 22, 2021

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
10:06

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
08:53

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

Area of Science:

  • Biochemistry
  • Genetics
  • Molecular Biology

Background:

  • Duchenne muscular dystrophy (DMD) is a severe genetic disorder caused by mutations in the dystrophin gene.
  • Antisense oligonucleotide (AO)-mediated exon skipping is a promising molecular therapy for DMD, aiming to restore dystrophin production.
  • Efficient and specific AO identification is crucial for the clinical success of this therapeutic approach.

Purpose of the Study:

  • To establish and validate an in vitro splicing system for optimizing antisense oligonucleotides (AOs) for Duchenne muscular dystrophy (DMD) therapy.
  • To describe the systematic process of identifying the most effective AO for inducing targeted exon skipping in DMD myocytes.
  • To demonstrate the utility of the developed system for optimizing AOs against various exons within the DMD gene.

Main Methods:

  • Primary cell culture of Duchenne muscular dystrophy (DMD) myocytes from biopsy samples.
  • Design and synthesis of a series of antisense oligonucleotides (AOs) targeting specific exons of the DMD gene.
  • Transfection of AOs into cultured DMD myocytes, followed by mRNA analysis using reverse transcription-PCR to assess exon skipping efficiency.

Main Results:

  • A systematic screening process using cultured DMD myocytes successfully identified optimized antisense oligonucleotides (AOs).
  • The in vitro splicing system allowed for the evaluation and comparison of AO efficacy in inducing specific exon skipping.
  • The developed methodology proved reliable for optimizing AOs against multiple target exons in the DMD gene.

Conclusions:

  • The established in vitro splicing system provides a simple and reliable method for identifying optimized antisense oligonucleotides (AOs).
  • This approach is essential for advancing molecular therapies for Duchenne muscular dystrophy (DMD) by ensuring the selection of highly effective AOs.
  • The validated methods facilitate the development of targeted exon skipping therapies for a broad range of DMD-causing mutations.