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Related Experiment Videos

Human apolipoprotein B gene intestinal control region.

T J Antes1, S A Goodart, W Chen

  • 1Research Institute, Palo Alto Medical Foundation, Palo Alto, California 94301, USA.

Biochemistry
|June 8, 2001
PubMed
Summary

Researchers identified novel regulatory elements in the apolipoprotein B (apoB) gene

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Area of Science:

  • Molecular Biology
  • Genetics
  • Gene Regulation

Background:

  • A previously identified 315 bp enhancer upstream of the human apolipoprotein B (apoB) gene directs high-level transgene expression in mice.
  • The distant apoB intestinal control region (ICR) contains regulatory elements crucial for intestinal apoB expression.

Purpose of the Study:

  • To further analyze the apoB ICR by examining segments near the 315 bp intestinal enhancer (315 IE).
  • To identify and characterize novel regulatory elements within the apoB ICR responsible for intestinal expression.

Main Methods:

  • DNaseI hypersensitivity (DH) studies to map regulatory regions.
  • Transient transfection assays using CaCo-2 cells to assess enhancer activity.
  • Gel retardation experiments to identify transcription factor binding sites.
  • Transgenic mouse models to evaluate in vivo gene expression.

Main Results:

  • Three new DH sites (DH2, DH3, DH4) were identified within the apoB ICR.
  • A 485 bp segment (485 IE) containing DH2 showed strong intestinal enhancer activity and drove transgene expression in mouse intestines.
  • An HNF-4/ARP-1 binding site was found within the 485 IE.
  • A 1031 bp segment (1031 IE) downstream of the 315 IE drove transgene expression but lacked enhancer activity.
  • All three identified ICR components exhibited similar apoB mRNA expression patterns in the intestine.
  • Liver-specific elements did not influence intestinal apoB transgene expression.

Conclusions:

  • The apoB ICR comprises multiple regulatory elements, including enhancers and other functional segments, that control intestinal apoB expression.
  • The 485 IE and 1031 IE represent novel functional components of the apoB ICR.
  • These findings enhance our understanding of the complex transcriptional regulation of the human apoB gene in the intestine.

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