Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

RNA Interference01:23

RNA Interference

RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
RNA Interference01:23

RNA Interference

RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
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...
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...
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,...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Phage portal proteins counteract stringent-response-mediated restriction.

Nature communications·2026
Same author

Lifestyle Intervention During Pregnancy Is Associated With Favourable Changes of Vitronectin in Women With Obesity: Results From the Randomised LiP Trial.

Clinical obesity·2026
Same author

Diabetes in pregnancy attenuates the sFlt-1/PlGF ratio for prediction of preeclampsia.

Pregnancy hypertension·2026
Same author

Prediction of term preeclampsia using angiogenic biomarkers at 24-29 weeks gestation.

Pregnancy hypertension·2026
Same author

Impact of maternal fasting blood glucose in pregnancy on body composition, anthropometric, and metabolic outcomes in newborns and 3-Year-Old offspring: results from the lifestyle in pregnancy and offspring study.

BMC pregnancy and childbirth·2026
Same author

Study protocol for a 15-year follow-up of a randomized controlled trial on lifestyle intervention in pregnancy: assessing long-term effects on body composition, metabolic traits, and mental health in mothers and offspring.

Trials·2026

Related Experiment Video

Updated: Jun 26, 2026

Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis
09:04

Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis

Published on: July 26, 2018

RNA decay by messenger RNA interferases.

Mikkel Christensen-Dalsgaard1, Martin Overgaard, Kristoffer Skovbo Winther

  • 1Institute for Cell and Molecular Biosciences, Medical School, University of Newcastle, Newcastle, United Kingdom.

Methods in Enzymology
|January 24, 2009
PubMed
Summary

This study details methods for producing and analyzing mRNA-cleaving enzymes like RelE from Escherichia coli. These enzymes, part of toxin-antitoxin systems, inhibit bacterial growth by stopping translation.

More Related Videos

Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing (RIPiT-Seq)
09:26

Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing (RIPiT-Seq)

Published on: July 10, 2019

Measurement of mRNA Decay Rates in Saccharomyces cerevisiae Using rpb1-1 Strains
12:21

Measurement of mRNA Decay Rates in Saccharomyces cerevisiae Using rpb1-1 Strains

Published on: December 13, 2014

Related Experiment Videos

Last Updated: Jun 26, 2026

Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis
09:04

Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis

Published on: July 26, 2018

Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing (RIPiT-Seq)
09:26

Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing (RIPiT-Seq)

Published on: July 10, 2019

Measurement of mRNA Decay Rates in Saccharomyces cerevisiae Using rpb1-1 Strains
12:21

Measurement of mRNA Decay Rates in Saccharomyces cerevisiae Using rpb1-1 Strains

Published on: December 13, 2014

Area of Science:

  • Molecular Biology
  • Bacteriology
  • Genetics

Background:

  • Toxin-antitoxin (TA) systems, including relBE and mazEF, are abundant in bacteria.
  • These systems encode mRNA-cleaving enzymes that inhibit translation and induce bacteriostasis.
  • Understanding these enzymes is crucial for controlling bacterial growth.

Purpose of the Study:

  • To describe protocols for overproduction, purification, and analysis of mRNA-cleaving enzymes.
  • To provide methods for studying RelE of Escherichia coli and its antitoxin RelB.
  • To present plasmid vectors for detailed cleavage site analysis in model mRNAs.

Main Methods:

  • Overproduction and purification of mRNA-cleaving enzymes.
  • Analysis of enzyme activity on model mRNAs.
  • Utilizing specific plasmid vectors for cleavage site mapping.

Main Results:

  • Established protocols for obtaining functional RelE and RelB proteins.
  • Demonstrated the utility of developed plasmid vectors for site-specific analysis.
  • Provided a framework for studying mRNA cleavage mechanisms in TA systems.

Conclusions:

  • The described protocols facilitate in-depth study of bacterial mRNA-cleaving enzymes.
  • This work aids in understanding the bacteriostatic mechanisms of TA systems.
  • The developed tools are valuable for research on bacterial gene regulation and antibiotic development.