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Related Concept Videos

RNA Stability01:53

RNA Stability

Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
RNA Stability01:53

RNA Stability

Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
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...
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
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...

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Defining the Program of Maternal mRNA Translation during In vitro Maturation using a Single Oocyte Reporter Assay
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An in vitro assay to study regulated mRNA stability.

D T Fritz1, L P Ford, J Wilusz

  • 1Department of Microbiology and Molecular Genetics, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA.

Science'S STKE : Signal Transduction Knowledge Environment
|December 26, 2001
PubMed
Summary

Researchers developed an in vitro mRNA turnover assay to study mRNA deadenylation and degradation. This system accurately mimics cellular processes and allows for the investigation of regulatory factors, including AU-rich elements (AREs).

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Last Updated: Jul 24, 2026

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Published on: June 16, 2021

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Posttranscriptional regulation of messenger RNA (mRNA) is crucial for cellular processes.
  • The complexity of mRNA regulation in mammalian cells necessitates simplified experimental systems.
  • Understanding mRNA turnover, including deadenylation and degradation, is key to deciphering cellular control mechanisms.

Purpose of the Study:

  • To develop and validate an in vitro mRNA turnover assay for studying mRNA deadenylation and degradation.
  • To investigate the factors influencing mRNA decay and regulation.
  • To provide a versatile system for analyzing specific RNA sequence activities and regulatory elements.

Main Methods:

  • Utilized an S100 HeLa cell extract and in vitro transcribed RNAs.
  • Developed an assay to mimic in vivo mRNA turnover end products.
  • Introduced synthetic RNA molecules, including those with AU-rich elements (AREs) from TNF-alpha and GM-CSF, to test sequence-specific activities.

Main Results:

  • The in vitro system accurately replicates mRNA deadenylation and decay, mirroring in vivo observations.
  • The assay demonstrated processive deadenylation and decay of capped and polyadenylated RNA without non-specific nuclease activity.
  • Incorporation of AREs from TNF-alpha and GM-CSF led to increased deadenylation and decay rates, consistent with in vivo findings.

Conclusions:

  • The established in vitro mRNA turnover assay is a reliable tool for studying mRNA deadenylation and degradation in mammalian cells.
  • The system allows for the detailed investigation of regulatory factors, such as AREs, impacting mRNA stability.
  • This adaptable assay holds potential for examining diverse mRNA regulatory events.