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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...

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

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...

Non-LTR Retrotransposons

Non-LTR Retrotransposons

As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...

Pre-mRNA Processing: RNA Splicing

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...

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Related Experiment Video

Updated: Jun 9, 2026

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models

Published on: December 9, 2016

Apparent non-canonical trans-splicing is generated by reverse transcriptase in vitro.

Jonathan Houseley¹, David Tollervey

¹Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, United Kingdom. jon.houseley@bbsrc.ac.uk

|September 1, 2010

Summary

This summary is machine-generated.

Most reported trans-splicing in mammals may be an artifact of reverse transcriptase template switching during cDNA synthesis. This study demonstrates template switching can create non-canonical splicing events, offering a diagnostic method to identify these artifacts.

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Substrate Generation for Endonucleases of CRISPR/Cas Systems

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Substrate Generation for Endonucleases of CRISPR/Cas Systems

Substrate Generation for Endonucleases of CRISPR/Cas Systems

Published on: September 8, 2012

Area of Science:

Molecular Biology
Genomics
RNA Biology

Background:

Trans-splicing, the joining of separate RNA molecules in vivo, is known in lower eukaryotes but was thought absent in metazoans.
Bioinformatic analyses suggested widespread trans-splicing in mammals, but these transcripts often lacked canonical splice sites, raising authenticity concerns.
Reverse transcriptase enzymes' template switching ability during transcription can generate artifacts resembling trans-spliced sequences.

Purpose of the Study:

To investigate the role of reverse transcriptase template switching in generating non-canonical RNA splicing events.
To develop an in vitro system for analyzing template switching during reverse transcription.
To differentiate true trans-splicing from experimental artifacts.

Main Methods:

Development of an in vitro system to analyze reverse transcriptase template switching.
Utilizing highly purified RNA substrates for reproducible experiments.
Comparative analysis of template switching products across different reverse transcriptases.

Main Results:

Demonstrated reproducible occurrence of apparent trans-splicing between two RNA molecules in vitro.
Detected other non-canonical splicing events, including exon shuffling and sense-antisense fusions.
Identified that sense-antisense fusions produce apparent antisense non-coding RNAs, which are reportedly abundant in humans.

Conclusions:

Proposed that most reported non-canonical splicing in metazoans results from reverse transcriptase template switching during cDNA preparation.
Showed that template switching products vary between different reverse transcriptases.
Established a diagnostic method to identify experimental artifacts arising from template switching.