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

Kristina Vintersten, Claudio Monetti, Marina Gertsenstein, Puzheng Zhang, Lajos Laszlo, Steffen Biechele, Andras Nagy

Research output: Contribution to journalArticleResearchpeer-review

246 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)241-246
Number of pages6
Journalgenesis: The Journal of Genetics and Development
Volume40
Issue number4
DOIs
Publication statusPublished - Dec 2004
Externally publishedYes

Keywords

  • Conditional gene expression
  • Cre recombinase
  • DsRed
  • lac Z
  • loxP
  • Red fluorescent protein
  • RFP
  • Tetraploid embryo complementation
  • Transgenic mice

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