Related Experiment Video

Updated: Jun 1, 2026

Nasal Potential Difference to Quantify Trans-epithelial Ion Transport in Mice
08:55

Nasal Potential Difference to Quantify Trans-epithelial Ion Transport in Mice

Published on: July 4, 2018

Nonsense-mediated mRNA decay and cystic fibrosis

Liat Linde1, Batsheva Kerem

  • 1Department of Genetics, The Life Sciences Institute, The Hebrew University, 91904, Jerusalem, Israel. liat.linde@mail.huji.ac.il

Methods in Molecular Biology (Clifton, N.J.)
|May 20, 2011
PubMed

Insights

This study details protocols for analyzing CFTR nonsense transcripts and investigating the role of nonsense-mediated mRNA decay (NMD) in readthrough treatment response for genetic diseases. Understanding NMD

Area of Science:

  • Molecular Biology
  • Genetics
  • Pharmacology

Background:

  • Premature termination codons (PTCs) are present in approximately one-third of genetic disease alleles.
  • Therapeutic strategies for PTCs focus on promoting translational readthrough to restore functional protein synthesis.
  • Variability in readthrough treatment response is influenced by PTC context, drug composition, and transcript levels.

Purpose of the Study:

  • To provide comprehensive protocols for analyzing CFTR nonsense transcript levels.
  • To investigate the role of nonsense-mediated mRNA decay (NMD) in modulating the response to readthrough therapy.
  • To establish methods for assessing CFTR function in the context of NMD and readthrough.

Main Methods:

  • Inhibition of the nonsense-mediated mRNA decay (NMD) pathway.
  • Quantification of CFTR transcripts containing premature termination codons (PTCs).
  • Measurement of physiologic NMD substrates and CFTR protein function.

Main Results:

  • Established protocols for quantifying PTC-bearing transcripts and assessing NMD activity.
  • Demonstrated the feasibility of analyzing the impact of NMD on readthrough treatment efficacy.
  • Provided a framework for understanding transcript-level regulation in therapeutic responses.

Conclusions:

  • Nonsense-mediated mRNA decay (NMD) is a key regulator of PTC-bearing transcript levels.
  • Understanding NMD's role is crucial for optimizing readthrough therapies for genetic diseases.
  • These protocols facilitate further research into personalized medicine approaches for PTC-related disorders.

More Related Videos

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells08:53

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells

Published on: May 16, 2017

Related Concept Videos

Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
Nuclear Export of mRNA02:31

Nuclear Export of mRNA

Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
Cystic Fibrosis: Pathogenesis01:23

Cystic Fibrosis: Pathogenesis

Cystic fibrosis (CF), an autosomal recessive disorder, significantly affects the function of exocrine glands. This genetically inherited disease is characterized by the production of thick and sticky mucus, which can severely affect various organs and systems in the body.
CF is primarily caused by a genetic mutation in a chromosome 7 gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR) protein. The most common gene mutation leading to CF is the ΔF508 mutation, but...
mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability