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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...

Alternative RNA Splicing

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 Splicing

Alternative RNA Splicing

Pre-mRNA Processing: RNA Splicing

Pre-mRNA Processing: RNA Splicing

Chromatin Structure Regulates pre-mRNA Processing

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...

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Related Experiment Video

Updated: Dec 25, 2025

Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts

Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts

Published on: May 11, 2018

Alpha-l-Locked Nucleic Acid-Modified Antisense Oligonucleotides Induce Efficient Splice Modulation In Vitro.

Prithi Raguraman^1,2, Tao Wang^1,2, Lixia Ma³

¹Centre for Molecular Medicine and Innovative Therapeutics, Murdoch University, Perth 6150 Australia.

International Journal of Molecular Sciences

|April 5, 2020

Summary

Alpha-l-Locked nucleic acid (α-l-LNA) antisense oligonucleotides (AOs) effectively induce Dmd exon-23 skipping in mdx mouse cells. Shorter α-l-LNA AOs demonstrate comparable or improved efficacy and low cytotoxicity compared to longer AOs.

Keywords:

DMD antisense oligonucleotides locked nucleic acids α-l-LNA

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Last Updated: Dec 25, 2025

Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts

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Using the E1A Minigene Tool to Study mRNA Splicing Changes

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Published on: April 22, 2021

Area of Science:

Oligonucleotide therapeutics
Molecular biology
Gene therapy

Background:

Locked nucleic acids (LNA) offer enhanced biophysical properties for oligonucleotide therapeutics.
Alpha-l-Locked nucleic acid (α-l-LNA) is a stereoisomer of LNA with potential for stable antisense oligonucleotides (AOs).
Antisense oligonucleotides can modulate gene splicing, offering therapeutic potential for genetic disorders.

Purpose of the Study:

To investigate the efficacy of α-l-LNA modified AOs in modulating gene splicing.
To assess the ability of α-l-LNA AOs to induce Dmd exon-23 skipping in vitro.
To evaluate the impact of AO size on splicing modulation efficacy and cytotoxicity.

Main Methods:

Synthesis of α-l-LNA modified 2'-O-methyl phosphorothioate (2'-OMePS) AOs of varying lengths (20mer, 18mer, 16mer).
In vitro evaluation of AO efficacy in inducing Dmd exon-23 skipping in mdx mouse myoblasts.
Comparison of truncated α-l-LNA AOs with fully modified 2'-OMePS control AOs.
Assessment of cytotoxicity of the evaluated AOs.

Main Results:

α-l-LNA modified AOs successfully induced Dmd exon-23 skipping in mdx mouse myoblasts.
Truncated 18mer and 16mer α-l-LNA AO variants showed comparable or slightly improved exon-skipping efficacy versus the control.
The truncated α-l-LNA AOs exhibited low cytotoxicity.
This study is the first to report the use of α-l-LNA modified AOs for splice modulation.

Conclusions:

α-l-LNA modified AOs are effective tools for splice modulation.
Truncated α-l-LNA AOs offer a promising strategy for developing stable and efficient antisense oligonucleotide therapeutics.
Further research into α-l-LNA AOs could expand their therapeutic applications, particularly in treating genetic disorders through splicing modulation.