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Related Experiment Videos

Codon-improved Cre recombinase (iCre) expression in the mouse.

D R Shimshek1, J Kim, M R Hübner

  • 1Department of Molecular Neuroscience, Max-Planck Institute for Medical Research, Heidelberg, Germany.

Genesis (New York, N.Y. : 2000)
|February 9, 2002
PubMed
Summary
This summary is machine-generated.

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We optimized Cre recombinase expression in mammalian cells and mice using mammalian codon usage and reduced CpG content, creating an improved Cre (iCre) for genetic experiments.

Area of Science:

  • Molecular Biology
  • Genetics
  • Mammalian Cell Culture

Background:

  • Cre recombinase is crucial for genetic engineering but its expression can be inefficient in mammalian systems.
  • Prokaryotic codon usage and high CpG content in the Cre gene can lead to poor expression and epigenetic silencing in mammals.

Purpose of the Study:

  • To enhance Cre recombinase expression and functionality in mammalian cells and transgenic mice.
  • To engineer an improved Cre (iCre) gene by optimizing codon usage and reducing CpG dinucleotides.

Main Methods:

  • Applied mammalian codon optimization to the Cre recombinase gene.
  • Reduced CpG content in the coding sequence to minimize epigenetic silencing.
  • Performed immunoblot and functional analyses to assess iCre expression and activity.

Related Experiment Videos

  • Generated transgenic mice expressing iCre and iCre fused to estrogen receptor domains.

    • Main Results:

    • Significantly improved Cre expression and function in three mammalian cell lines.
    • Successfully generated transgenic mice with high frequency of iCre expression.
    • Demonstrated efficient loxP-mediated DNA recombination in iCre-expressing cells in mice.
    • Observed limited Cre activity upon tamoxifen induction in tamoxifen-inducible Cre fusion models.

    Conclusions:

    • The engineered iCre facilitates efficient genetic experiments in mice.
    • Further optimization is needed for temporal control of Cre fusion proteins, particularly in the brain.