Quality control of astrocyte-directed Cre transgenic mice: the benefits of a direct link between loss of gene expression and reporter activation

Glia. 2009 Apr 15;57(6):680-92. doi: 10.1002/glia.20796.

Abstract

Cre recombinase activity for cell-type restricted deletion of floxed target genes (i.e., flanked by Cre recognition loxP-sites) is often measured by separate matings with recombination-activated reporter gene mice. Using a floxed Gja1 (Cx43) allele, we demonstrate the benefits of a direct link between reporter gene expression and target gene deletion to overcome critical limitations of the Cre/loxP system. The widely used human glial fibrillary acidic protein (hGFAP)-Cre transgene exhibits variable recombination activity and requires postexperimental validation. Such quality control is essential to correlate the extent of Cre-mediated Gja1 ablation with phenotypical alterations and to maintain the activity status of hGFAP-Cre in transgenic mouse colonies. We present several strategies to control for the fidelity of hGFAP-Cre mediated recombination. (c) 2008 Wiley-Liss, Inc.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Astrocytes / physiology
  • Connexins / genetics
  • Female
  • Gene Deletion
  • Gene Expression*
  • Gene Transfer Techniques*
  • Genes, Reporter*
  • Glial Fibrillary Acidic Protein / genetics
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • Male
  • Methylation
  • Mice
  • Mice, Transgenic
  • Promoter Regions, Genetic / physiology
  • Quality Control

Substances

  • Connexins
  • Glial Fibrillary Acidic Protein