Related Experiment Video
Updated: Dec 23, 2025
11:48
Detection of Alternative Splicing During Epithelial-Mesenchymal Transition
Published on: October 9, 2014
13.2K
Subphysiological temperature induces pervasive alternative splicing in Chinese hamster ovary cells
Ioanna Tzani1, Craig Monger1, Krishna Motheramgari1,2
1National Institute for Bioprocessing Research and Training, Dublin, Ireland.
Biotechnology and Bioengineering
|April 30, 2020
Summary
This study reveals significant alternative messenger RNA (mRNA) splicing events in Chinese hamster ovary (CHO) cells during biopharmaceutical production. These post-transcriptional changes offer new insights into cellular responses to temperature shifts.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- RNA sequencing (RNASeq) is common for analyzing gene expression in Chinese hamster ovary (CHO) cells.
- Alternative messenger RNA (mRNA) splicing in CHO cells remains understudied despite its potential impact on bioprocesses.
Purpose of the Study:
- To investigate alternative splicing events in a monoclonal antibody (mAb)-producing CHO K1 cell line under temperature shift conditions.
- To explore the role of post-transcriptional regulation in CHO cell biopharmaceutical production.
Main Methods:
- Utilized RNA sequencing (RNASeq) for transcriptomic analysis of CHO K1 cells.
- Applied differential splicing analysis to identify alternative mRNA splicing events.
- Validated key splicing events using quantitative polymerase chain reaction (qPCR).
Main Results:
- Identified over 2,465 instances of differential splicing 24 hours after a temperature reduction.
- Found 1,197 alternative splicing events in genes without significant changes in expression levels.
- Validated exon skipping and mutually exclusive splicing in genes related to temperature response and mitochondrial function.
Conclusions:
- Alternative splicing analysis provides a deeper understanding of post-transcriptional regulation in CHO cells.
- Identified splicing events are linked to cellular responses to temperature changes and mitochondrial function.
- Further research is needed to elucidate the phenotypic impact of these alternative splicing events.
More Related Videos
Related Concept Videos
02:18Alternative RNA Splicing
24.4K
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...
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...
24.4K
02:18Alternative RNA Splicing
4.6K
4.6K
01:32RNA Splicing
60.1K
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...
60.1K
02:28Leaky Scanning
5.5K
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.5K
02:27Background and Environment Affect Phenotype
7.3K
Although the genetic makeup of an organism plays a major role in determining the phenotype, there are also several environmental factors, such as temperature, oxygen availability, presence of mutagens, that can alter an organism’s phenotype.
An example of how genetic background affects phenotype can be seen in horses. The Extension gene in horses is responsible for their coat color. A wild-type gene (EE) produces black pigment in the coat, while a mutant gene (ee) produces red pigment. A...
An example of how genetic background affects phenotype can be seen in horses. The Extension gene in horses is responsible for their coat color. A wild-type gene (EE) produces black pigment in the coat, while a mutant gene (ee) produces red pigment. A...
7.3K
02:32Exon Recombination
4.0K
The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes.
Exon shuffling follows “splice frame rules.” Each exon...
Exon shuffling follows “splice frame rules.” Each exon...
4.0K