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Mouse in red: Red fluorescent protein expression in mouse ES cells, embryos, and adult animals
Author(s) -
Vintersten Kristina,
Monetti Claudio,
Gertsenstein Marina,
Zhang Puzheng,
Laszlo Lajos,
Biechele Steffen,
Nagy Andras
Publication year - 2004
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.20095
Subject(s) - green fluorescent protein , biology , recombinase , transgene , microbiology and biotechnology , embryo , genetically modified mouse , cell culture , cre recombinase , gene , genetics , recombination
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. genesis 40:241–246, 2004. © 2004 Wiley‐Liss, Inc.