C57BL/6-Rabgef1^tm3.1Glli/J

Stock number: 034063
Product name: Rabgef1^fl
Research areas: Cell Biology Research, Developmental Biology Research, Immunology, Inflammation and Autoimmunity Research

Description

Rabgef1^fl/fl mice possess loxP sites flanking exon 2 of the RAB guanine nucleotide exchange factor (GEF) 1 (Rabgef1) gene. These floxed mice are useful when studying the role of Rabgef1 in epidermal barrier function.

Detailed description

Rabgef1^fl/fl mice contain loxP sites flanking exon 2 of the RAB guanine nucleotide exchange factor (GEF) 1 (Rabgef1) gene. The gene product, RABGEF1, is a guanine nucleotide exchange factor for Rab5 and is involved in endocytic membrane fusion and membrane trafficking of recycling endosomes. RABGEF1 has also been shown to be a negative regulator of mast cell activation and Ras-mediated signaling pathways. Mice that are homozygous for this floxed allele are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 2 deleted in the Cre-expressing tissues.

For example, when bred to the mice with K14-Cre mice with cre expression in skin, the resulting Rabgef1^K-KO mice exhibit spontaneous skin inflammation with features of human atopic dermatitis. They exhibit impaired epidermal barrier function and an allergic cutaneous and systemic phenotype. RABGEF1-deficient keratinocytes exhibit a dysfunction in IL-1R/MYD88/NF-κB signaling pathway and MYD88-dependent abnormalities in expression of structural proteins that contribute to skin barrier function.

Development

The Rabgef1^fl targeting vector was designed to insert a loxP site upstream of exon 2 of the RAB guanine nucleotide exchange factor (GEF) 1 (Rabgef1) gene, and a second loxP site downstream of exon 2, followed by a frt-flanked neomycin selection cassette. The construct was electroporated into C57BL/6N embryonic stem (ES) cells and correctly targeted ES cells were injected into recipient blastocysts. Chimeras were bred with mice expressing Flp recombinase on a C57BL/6J background to remove the neo cassette, and the progeny were then crossed to remove the FLP-expressing transgene. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).



In 2019 a 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 43 markers throughout the genome suggested a C57BL/6 genetic background, 1 of 5 markers, on chromosome 3, was segregating with an unknown source.

Allele

Symbol: Rabgef1^tm3.1Glli
Name: targeted mutation 3.1, Stephen J Galli
MGI accession ID: MGI:6281617
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Synonyms: Rabgef1^fl
Marker symbol: Rabgef1
Marker name: RAB guanine nucleotide exchange factor (GEF) 1
Marker synonyms: RABEX5; rabex-5; RAP1; Ras negative regulator Rabex-5
Chromosome: 5
Strain of origin: C57BL/6N
Molecular note: Exon 2 was floxed. Flp-mediated recombination removed the neomycin seletion cassette.