Silencing inhibits Cre-mediated recombination of the Z/AP and Z/EG reporters in adult cells

PLoS One. 2009;4(5):e5435. doi: 10.1371/journal.pone.0005435. Epub 2009 May 5.

Abstract

Background: The Cre-loxP system has been used to enable tissue specific activation, inactivation and mutation of many genes in vivo and has thereby greatly facilitated the genetic dissection of several cellular and developmental processes. In such studies, Cre-reporter strains, which carry a Cre-activated marker gene, are frequently utilized to validate the expression profile of Cre transgenes, to act as a surrogate marker for excision of a second allele, and to irreversibly label cells for lineage tracing experiments.

Principal findings: We have studied three commonly used Cre-reporter strains, Z/AP, Z/EG and R26R-EYFP and have demonstrated that although each reporter can be reliably activated by Cre during early development, exposure to Cre in adult hematopoietic cells results in a much lower frequency of marker-positive cells in the Z/AP or Z/EG strains than in the R26R-EYFP strain. In marker negative cells derived from the Z/AP and Z/EG strains, the transgenic promoter is methylated and Cre-mediated recombination of the locus is inhibited.

Conclusions: These results show that the efficiency of Cre-mediated recombination is not only dependent on the genomic context of a given loxP-flanked sequence, but also on stochastic epigenetic mechanisms underlying transgene variegation. Furthermore, our data highlights the potential shortcomings of utilizing the Z/AP and Z/EG reporters as surrogate markers of excision or in lineage tracing experiments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult Stem Cells
  • Animals
  • Epigenesis, Genetic
  • Gene Silencing*
  • Genes, Reporter / genetics*
  • Genetic Markers
  • Hematopoietic Stem Cells / metabolism*
  • Integrases / metabolism*
  • Mice
  • Mice, Transgenic
  • Transgenes

Substances

  • Genetic Markers
  • Cre recombinase
  • Integrases