Germ-line recombination activity of the widely used hGFAP-Cre and nestin-Cre transgenes

PLoS One. 2013 Dec 9;8(12):e82818. doi: 10.1371/journal.pone.0082818. eCollection 2013.

Abstract

Herein we demonstrate with PCR, immunodetection and reporter gene approaches that the widely used human Glial Fibrillary Acidic Protein (hGFAP)-Cre transgene exhibits spontaneous germ-line recombination activity in leading to deletion in brain, heart and tail tissue with high frequency. The ectopic activity of hGFAP-Cre requires a rigorous control. We likewise observed that a second widely used nestin-Cre transgene shows germ-line deletion. Here we describe procedures to identify mice with germ-line recombination mediated by the hGFAP-Cre and nestin-Cre transgenes. Such control is essential to avoid pleiotropic effects due to germ-line deletion of loxP-flanked target genes and to maintain the CNS-restricted deletion status in transgenic mouse colonies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cerebral Cortex / metabolism
  • Connexin 43 / genetics
  • Female
  • Gene Deletion
  • Gene Knockout Techniques
  • Gene Order
  • Germ Cells / metabolism*
  • Glial Fibrillary Acidic Protein / genetics*
  • Glial Fibrillary Acidic Protein / metabolism
  • Hippocampus / metabolism
  • Homologous Recombination*
  • Humans
  • Immunohistochemistry
  • Integrases / genetics*
  • Integrases / metabolism*
  • Male
  • Mice
  • Mice, Knockout
  • Mice, Transgenic
  • Myocardium / metabolism
  • Nestin / genetics*
  • Transgenes

Substances

  • Connexin 43
  • Glial Fibrillary Acidic Protein
  • Nestin
  • Cre recombinase
  • Integrases

Grants and funding

This work was supported by grants of the German Research Foundation SFB/TR3, C1 and C9; SPP1172 SE 774/3 (to CS); SFB/TR3, N01 and C9; SPP1172 TH1350/1-1 (to MT) and of the European Community FP7-202167 (to CS and MT). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.