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Cre transgene results in global attenuation of the cAMP/PKA pathway

L Gangoda1, M Doerflinger, Y Y Lee

  • 1Department of Biochemistry, La Trobe Institute of Molecular Science, La Trobe University, Kingsbury Drive, Bundoora, Victoria, Australia 3086.

Cell Death & Disease
|August 10, 2012
PubMed

Insights

Cre recombinase expression in transgenic models causes unintended changes in protein kinase A (PKA) activity and gene expression, impacting experimental results. These findings may necessitate re-evaluation of data from conventional Cre-mediated gene deletion studies.

Area of Science:

  • Molecular Biology
  • Genetics
  • Immunology

Background:

  • Cre recombinase is widely used in transgenic mice to delete floxed alleles for gene function studies.
  • Understanding off-target effects of Cre expression is crucial for accurate interpretation of experimental data.

Purpose of the Study:

  • To investigate the impact of Cre recombinase expression on protein kinase A (PKA) signaling pathways.
  • To determine if Cre expression affects cellular processes like cell death and cytokine secretion.

Main Methods:

  • Utilized cultured cell lines and transgenic mouse models expressing Cre recombinase.
  • Analyzed PKA target phosphorylation, gene expression profiles, and cytokine secretion (e.g., IL-6).
  • Investigated the role of PKA inhibitors (PKI) in mediating Cre-induced effects.

Main Results:

  • Cre expression led to global alterations in PKA target phosphorylation in both cell lines and mice.
  • Observed changes in gene expression profiles and increased secretion of cytokines like IL-6.
  • These effects were dependent on Cre's recombinase activity and linked to upregulated PKI.

Conclusions:

  • Cre expression induces significant, unintended molecular and cellular changes.
  • These alterations can be attributed to PKA pathway dysregulation and PKI upregulation.
  • Findings may explain observed cytotoxicity and phenotypes in Cre-based transgenic systems, influencing data interpretation.

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