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Related Concept Videos

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

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A Suppressor Screen for the Characterization of Genetic Links Regulating Chronological Lifespan in Saccharomyces cerevisiae
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Validation of suitable internal control genes for expression studies in aging.

M Zampieri1, F Ciccarone, T Guastafierro

  • 1Department of Cellular Biotechnologies and Hematology, II Faculty of Medicine and Surgery, Sapienza University of Rome, Viale Regina Elena 324, 00161 Rome, Italy.

Mechanisms of Ageing and Development
|December 30, 2009
PubMed
Summary

This study evaluated housekeeping genes for accurate gene expression normalization in human blood cells across different ages. Glucuronidase (GUSB) showed the most stable expression, making it ideal for aging research.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Immunology

Background:

  • Gene expression normalization commonly relies on housekeeping genes, assuming stable expression.
  • This assumption is increasingly challenged by evidence of variable expression in specific biological contexts.
  • Reference gene validation in aging research is underdeveloped, lacking defined standards.

Purpose of the Study:

  • To assess the expression stability of commonly used housekeeping genes in human peripheral blood mononuclear cells (PBMCs) across an aging spectrum.
  • To identify reliable reference genes for accurate gene expression analysis in aging studies.

Main Methods:

  • Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was employed.
  • Expression stability of five housekeeping genes (18s rRNA, ACTB, GAPDH, HPRT1, GUSB) was analyzed in PBMCs from donors aged 35-74 years.
  • Stability was quantified using coefficient of variation, geNorm, and NormFinder algorithms.

Main Results:

  • Glucuronidase (GUSB) exhibited the highest expression stability, identifying it as the most suitable reference gene.
  • 18s ribosomal RNA (rRNA) demonstrated the least stability, making it unsuitable for normalization in this context.
  • Aging was identified as a significant factor influencing the stability of 18s rRNA, hypoxanthine phosphoribosyltransferase 1 (HPRT1), and actin beta (ACTB) expression.

Conclusions:

  • GUSB is recommended as a reliable reference gene for normalizing gene expression in human PBMCs, particularly in aging studies.
  • 18s rRNA is not a suitable housekeeping gene for this cell type and age range.
  • Researchers must consider age as a critical variable when selecting reference genes for gene expression studies in PBMCs.