B6N;129S6-Eef1a1^tm1Tcrl/J

Stock number: 027588
Product name: EF1a^Crb3-GFP
Research areas: Research Tools

Description

EF1a^Crb3-GFP KI mice may be useful as a Cre mediated reporter for visualizing membrane and/or tissue dynamics in living tissues.

Detailed description

EF1a^Crb3-GFP KI mice contain a floxed STOP cassette and a crumbs family member 3 (Crb3)-enhanced green fluorescent protein (GFP) fusion protein (Crb3-GFP) in exon 1 of the eukaryotic translation elongation factor 1 alpha 1 (Eef1a1) gene, abolishing Eef1a1 expression. Crumbs3 is a transmembrane protein localized to the apical membranes of epithelial cells. Homozygous Crb3-GFP males are viable and fertile, while homozygous females are not viable. In the absence of Cre, reporter gene expression is prevented by the STOP sequence. After removal of the loxP-flanked STOP cassette via cre-mediated recombination, GFP labels apical membranes in the majority of epithelial cell types where Cre recombinase is expressed.

Development

The Crb3-GFP construct was designed with a loxP-flanked neomycin resistance (neo) cassette followed by a crumbs family member 3 (Crb3)-enhanced green fluorescent protein (GFP) fusion protein. The floxed-neo acts as a transcriptional STOP cassette. The construct was targeted to exon 1 of the eukaryotic translation elongation factor 1 alpha 1(Eef1a1) gene and was electroporated into 129S6/SvEvTac-derived SM-1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric males were bred to C57BL/6N. These mice were maintained on a mixed 129;C57BL/6N background. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ inbred mice (Stock No. 005304) for at least one generation.