Mouse in red: red fluorescent protein expression in mouse ES cells, embryos, and adult animals

Genesis. 2004 Dec;40(4):241-6. doi: 10.1002/gene.20095.

Abstract

Spectral variants of green fluorescent protein are widely used in live samples for a broad range of applications: from visualization of protein interactions, through following gene expression, to marking particular cells in complex tissues. Higher wavelength emissions (such as red) are preferred due to the lower background-autofluorescence in tissues (Miyawaka et al., Nat Cell Biol Suppl S1-7, 2003). Until now, however, red fluorescent proteins (RFP) have displayed toxicity in murine embryos, which has hampered its application in this model. Here we report strong expression of a recently developed RFP variant, DsRed.T3, in mouse ES cells, embryos, and adult mice. Our results show that the red fluorescent wavelength has a superior tissue penetrance compared with spectral variants of lower wavelength. Furthermore, we have generated an ES cell line and a corresponding transgenic mouse line in which red fluorescence is activated upon Cre excision. Finally, we introduced cell type-specifically expressed Cre transgenes into this Cre recombinase reporter cell line, and by using the tetraploid embryo complementation assay, we could directly verify the Cre recombinase specificity on ES cell-derived embryos/animals.

MeSH terms

  • Animals
  • Cells, Cultured
  • Electroporation
  • Embryo, Mammalian / metabolism*
  • Gene Expression
  • Genes, Reporter
  • Integrases / genetics
  • Lac Operon
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, Transgenic
  • Stem Cells / metabolism*
  • Transgenes
  • Viral Proteins / genetics

Substances

  • Luminescent Proteins
  • Viral Proteins
  • red fluorescent protein
  • Cre recombinase
  • Integrases