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Related Experiment Video

Updated: May 23, 2026

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Published on: July 28, 2017

Optimizing splice-switching oligomer sequences using 2'-O-methyl phosphorothioate chemistry.

Carl Adkin¹, Sue Fletcher, Steve D Wilton

¹Centre for Neuromuscular and Neurological Disorders, University of Western Australia, Perth, Australia.

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

|March 29, 2012

Summary

Researchers designed splice-switching oligomers to induce targeted dystrophin exon skipping. This approach analyzes exon sequences to create effective antisense compounds for potential therapeutic applications.

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Chemical Triphosphorylation of Oligonucleotides

Chemical Triphosphorylation of Oligonucleotides

Published on: June 2, 2022

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Last Updated: May 23, 2026

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Published on: July 28, 2017

Chemical Triphosphorylation of Oligonucleotides

Chemical Triphosphorylation of Oligonucleotides

Published on: June 2, 2022

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

Duchenne muscular dystrophy (DMD) is caused by mutations in the dystrophin gene.
Exon skipping is a therapeutic strategy to restore the dystrophin reading frame.
Developing effective splice-switching oligomers is crucial for this strategy.

Purpose of the Study:

To develop an empirical approach for designing splice-switching oligomers.
To induce targeted dystrophin exon skipping in myogenic cells.
To identify effective antisense compounds for potential clinical applications.

Main Methods:

Analysis of exon sequences for splice site motifs and enhancers.
Design and synthesis of 2'-O-methyl-modified oligomers on a phosphorothioate backbone (2OMeAOs).
Transfection of 2OMeAOs into cultured myogenic cells and analysis by semi-quantitative RT-PCR.

Main Results:

Demonstrated reproducible and dose-dependent induction of exon skipping.
Identified specific oligomers capable of modulating dystrophin pre-mRNA splicing.
Established a method for evaluating oligomer efficacy in vitro.

Conclusions:

The empirical design approach is effective for creating splice-switching oligomers.
Targeted exon skipping can be induced in myogenic cells using 2OMeAOs.
Further development is needed for clinical application, focusing on low-dosage efficacy and safety.