Mouse in red: red fluorescent protein expression in mouse ES cells, embryos, and adult animals.
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.
Vintersten
Kristina
K
Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
Monetti
Claudio
C
Gertsenstein
Marina
M
Zhang
Puzheng
P
Laszlo
Lajos
L
Biechele
Steffen
S
Nagy
Andras
A
D000818:Animals; D002478:Cells, Cultured; D018274:Electroporation; D004622:Embryo, Mammalian / Q000378:metabolism*; D015870:Gene Expression; D017930:Genes, Reporter; D019426:Integrases / Q000235:genetics; D007763:Lac Operon; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D013234:Stem Cells / Q000378:metabolism*; D019076:Transgenes; D014764:Viral Proteins / Q000235:genetics; D000097573:Red Fluorescent Protein
D019426:Integrases; D008164:Luminescent Proteins; D014764:Viral Proteins; D000097573:Red Fluorescent Protein; C045073:Cre recombinase