Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Nonsense-mediated mRNA Decay

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,...

mRNA Stability and Gene Expression

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

You might also read

Related Articles

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

Sort by

Same author

Uncovering the initial response: Intra-mitochondrial surveillance activates the UPR<sup>mt</sup>.

Molecular cell·2026

Same author

Probing direct interactions between nuclear proteins in cells with nxReLo.

Proceedings of the National Academy of Sciences of the United States of America·2025

Same author

Molecular insights into the Drosophila piRNA pathway via systematic ReLo protein interaction screening and structure prediction.

Nucleic acids research·2025

Same author

Recurrent innovation of protein-protein interactions in the Drosophila piRNA pathway.

The EMBO journal·2025

Same author

ReLo is a simple and rapid colocalization assay to identify and characterize direct protein-protein interactions.

Nature communications·2024

Same author

In Vitro Reconstitution of the Drosophila melanogaster CCR4-NOT Complex to Assay Deadenylation.

Methods in molecular biology (Clifton, N.J.)·2023

Search research articles

Related Experiment Video

Updated: May 2, 2026

Antibody-Free Assay for RNA Methyltransferase Activity Analysis

Antibody-Free Assay for RNA Methyltransferase Activity Analysis

Published on: July 9, 2019

Assaying mRNA deadenylation in vitro.

Mandy Jeske¹, Claudia Temme, Elmar Wahle

¹Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Kurt-Mothes-Strasse 3, 06120, Halle, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|March 5, 2014

Summary

This study details two in vitro methods for assaying mRNA deadenylation, the removal of poly(A) tails. These techniques enable the study of deadenylation enzymes and sequence-specific RNA degradation.

More Related Videos

An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity

An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity

Published on: October 8, 2018

Dot Blot Assay for Detecting Global N6-Methyladenosine RNA Modification Levels

Dot Blot Assay for Detecting Global N⁶-Methyladenosine RNA Modification Levels

Published on: February 6, 2026

Related Experiment Videos

Last Updated: May 2, 2026

Antibody-Free Assay for RNA Methyltransferase Activity Analysis

Antibody-Free Assay for RNA Methyltransferase Activity Analysis

Published on: July 9, 2019

An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity

An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity

Published on: October 8, 2018

Dot Blot Assay for Detecting Global N6-Methyladenosine RNA Modification Levels

Dot Blot Assay for Detecting Global N⁶-Methyladenosine RNA Modification Levels

Published on: February 6, 2026

Area of Science:

Molecular Biology
Biochemistry
RNA Metabolism

Background:

Deadenylation, the removal of the poly(A) tail from mRNA, is a critical step in RNA regulation.
Understanding deadenylation mechanisms is essential for comprehending gene expression control.

Purpose of the Study:

To present two novel in vitro assay methods for quantifying mRNA deadenylation.
To facilitate the study of enzymes involved in poly(A) tail shortening and RNA decay.

Main Methods:

Development of a rapid, quantitative assay for radiolabeled poly(A) degradation.
Establishment of a Drosophila embryo extract-based assay for sequence-specific, in vivo-directed deadenylation.

Main Results:

The first method offers a versatile tool adaptable to various deadenylation enzymes.
The second method demonstrates sequence-specific deadenylation in vitro, mirroring in vivo processes.

Conclusions:

The presented in vitro assays provide robust and adaptable methods for studying mRNA deadenylation.
These assays will aid in dissecting the enzymatic and sequence-specific aspects of RNA deadenylation and decay.