Comparative Analysis of the Efficiency and Specificity of myeloid-Cre Deleting Strains Using ROSA-EYFP Reporter Mice

J Immunol Methods. 2014 Jun;408:89-100. doi: 10.1016/j.jim.2014.05.009. Epub 2014 May 22.

Abstract

Since the first example of conditional gene targeting in mice in 1994, the use of Cre recombinase and loxP flanked sequences has become an invaluable technique to generate tissue and temporal specific gene knockouts. The number of mouse strains expressing floxed-gene sequences, and tissue-specific or temporal-specific Cre-recombinase that have been reported in the literature has grown exponentially. However, increased use of this technology has highlighted several problems that can impact the interpretation of any phenotype observed in these mouse models. In particular, accurate knowledge of the specificity of Cre expression in each strain is critical in order to make conclusions about the role of specific cell types in the phenotypes observed. Cre-mediated deletion specificity and efficiency have been described in many different ways in the literature, making direct comparisons between these Cre strains impossible. Here we report crossing thirteen different myeloid-Cre mouse strains to ROSA-EYFP reporter mice and assaying YFP expression in a variety of naïve unstimulated hematopoietic cells, in parallel. By focusing on myeloid subsets, we directly compare the relative efficiency and specificity of myeloid deletion in these strains under steady-state conditions.

Keywords: Cre-recombinase; Dendritic cell; Floxed; Macrophage; Neutrophil.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bacterial Proteins / biosynthesis*
  • Bacterial Proteins / genetics
  • Flow Cytometry
  • Gene Deletion*
  • Gene Targeting*
  • Genes, Reporter*
  • Genotype
  • Integrases / genetics*
  • Luminescent Proteins / biosynthesis*
  • Luminescent Proteins / genetics
  • Mice
  • Mice, 129 Strain
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Myeloid Cells / metabolism*
  • Phenotype
  • RNA, Untranslated / genetics*

Substances

  • Bacterial Proteins
  • Gt(ROSA)26Sor non-coding RNA, mouse
  • Luminescent Proteins
  • RNA, Untranslated
  • yellow fluorescent protein, Bacteria
  • Cre recombinase
  • Integrases