B6.129P2(C)-Arhgef11^tm1.1Mak/J

Stock number: 029252
Product name: PDZ-RhoGEF-knockout
Research areas: Diabetes and Obesity Research, Endocrine Deficiency Research, Internal/Organ Research, Metabolism Research, Research Tools

Description

This Arhgef11 (PDZ-RhoGEF) knockout strain is useful in studies of adipose tissue development, energy homeostasis, and resistance to diet induced obesity and type 2 diabetes.

Detailed description

The targeted Arhgef11 gene encodes PDZ-RhoGEF, a Rho GTPase that is involved in cell signaling, cell migration, chemotaxis, neurite outgrowth, as well as metabolic homeostasis. Mutations in this gene have been associated with obesity and type 2 diabetes. These knock-out mice carry a mutation in which exon 2 has been deleted via Cre-mediated recombination. No gene product (protein) is detected by Western blot analysis of MEFs isolated from homozygotes.

Mice that are homozygous for the targeted mutation are viable and fertile, but are smaller in size than wildtype controls. Homozygotes exhibit lower body fat index with reduced major fat depots (epididymal, knee, inguinal, and retroperitoneal white adipose tissue) when compared to controls, and a slight reduction in brown adipose tissue mass. Fewer mature adipocytes are found in white adipose. MEFs from mutant mice exhibit reduced cell proliferation. Homozygotes on a high fat diet do not develop obesity, glucose intolerance or insulin resistance. Total oxygen consumption is increased in homozygotes, and there is a trend to increased total activity. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6 genetic background.

Development

A FRT site flanked targeting vector containing a PGK-Neo selection cassette and a 3’ end loxP was utilized in the construction of this mutant. This selection cassette was inserted upstream of exon 2 of the targeted gene, and another loxP site was inserted downstream of exon 2. The construct was electroporated into 129P2/OlaHsd derived E14K embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were tested for germline transmission. Heterozygotes were crossed with B6.C-Tg(CMV-cre)1Cgn/J mice (Stock No. 006054) to delete the floxed exon 2 and PGK-neo cassette. The mice were then backcrossed to C57BL/6 for 6 generations. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6 genetic background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Arhgef11^tm1.1Mak
Name: targeted mutation 1.1, Tak Mak
MGI accession ID: MGI:5788572
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Arhgef11
Marker name: Rho guanine nucleotide exchange factor (GEF) 11
Chromosome: 3
Strain of origin: 129P2/OlaHsd
Molecular note: A loxP site, followed by an FRT site, a neomycin selection cassette, a second loxP site and an FRT site are inserted upstream of exon 2, and a third loxP site was inserted downstream of exon 2. Cre-mediated recombination removed exon 2. Western blot and immunoblot analysis confirms that no protein was expressed in mutant mice.