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Related Experiment Videos

A generic intron increases gene expression in transgenic mice.

T Choi1, M Huang, C Gorman

  • 1Department of Biology, Massachusetts Institute of Technology, Cambridge 02142.

Molecular and Cellular Biology
|June 1, 1991
PubMed
Summary

Adding a specific hybrid intron significantly boosts gene expression in transgenic mice. This splicing element, comprising adenovirus and immunoglobulin G sequences, enhances chloramphenicol acetyltransferase (CAT) activity, proving valuable for gene expression studies.

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Area of Science:

  • Molecular Biology
  • Gene Expression Regulation

Background:

  • Splicing plays a crucial role in gene expression regulation.
  • Understanding how introns influence transgene expression is vital for genetic studies.

Purpose of the Study:

  • To investigate the impact of a heterologous intron on gene expression in transgenic mice.
  • To compare the efficacy of a hybrid intron versus a simian virus 40 intron in enhancing transgene expression.

Main Methods:

  • Generated transgenic mice with a human histone H4 promoter linked to the chloramphenicol acetyltransferase (CAT) gene.
  • Incorporated a hybrid intron (adenovirus splice donor/immunoglobulin G splice acceptor) or a simian virus 40 intron into the transgene.
  • Assessed CAT activity across various tissues to quantify gene expression levels.

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Main Results:

  • Transgenes with the hybrid intron showed a 5- to 300-fold increase in CAT activity compared to those without an intron.
  • The hybrid intron enhanced expression broadly across tissues and increased the frequency of transgenics with significant CAT activity.
  • The hybrid intron was significantly more effective than the simian virus 40 intron in elevating transgene expression.

Conclusions:

  • The inclusion of a specific hybrid intron can substantially enhance gene expression in transgenic mice.
  • This hybrid intron offers a valuable tool for achieving high-level transgene expression, potentially improving the utility of cDNA constructs in research.