Unspecific DNA recombination in AdipoqCre-ERT2 - mediated knockout approaches in transgenic mice is sex-, age- and genotype-dependent

Adipocyte. 2020 Dec;9(1):1-6. doi: 10.1080/21623945.2019.1701394.

Abstract

Due to the epidemic rise of obesity prevalence, adipose tissue (AT) research is of major interest. Our aim was to study specificity of the most-common Cre/loxP approach for inducible gene manipulation of AT in mice (AdipoqCre-ERT2). We used mice with tamoxifen-sensitive Cre recombinase controlled by the adiponectin promoter (AdipoqCre-ERT2), which were crossed to a tdTomato reporter mouse to visualize the site of recombination on a single-cell resolution. Albeit tamoxifen induced tdTomato expression in this model, also non-stimulated background recombination ('Cre leakage') was detected in AT of untreated Adipoq-CreERT2xTDTO mice in vivo. Quantification of Cre leakage revealed age, sex and genotype as factors impacting on non-induced Cre recombination.

Keywords: Cre leakage; Cre recombinase; Cre/loxP; adiponectin promoter; adipose tissue; diabetes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / metabolism*
  • Age Factors
  • Animals
  • Cells, Cultured
  • DNA / genetics*
  • Genotype
  • Integrases / metabolism*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Transgenic
  • Receptors, Estrogen / metabolism*
  • Recombination, Genetic / genetics*
  • Sex Factors

Substances

  • Receptors, Estrogen
  • DNA
  • Cre recombinase
  • Integrases

Grants and funding

This work was supported by the Deutsche Forschungsgemeinschaft [209933838]; Deutsche Forschungsgemeinschaft [209933838].