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.
Dr. Tak Mak, University Health Network/Un of Toronto
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Arhgef11 | Rho guanine nucleotide exchange factor (GEF) 11 |
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.
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 Name | targeted mutation 1.1, Tak Mak |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | PDZ-RhoGEF KO |
Gene Symbol and Name | Arhgef11, Rho guanine nucleotide exchange factor (GEF) 11 |
Gene Synonym(s) | |
Strain of Origin | 129P2/OlaHsd |
Chromosome | 3 |
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. |
When maintaining a live colony, these mice can be bred as homozygotes.
When using the PDZ-RhoGEF-knockout mouse strain in a publication, please cite the originating article(s) and include JAX stock #029252 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Arhgef11<tm1.1Mak> |
Frozen Mouse Embryo | B6.129P2(C)-Arhgef11<tm1.1Mak>/J | $2595.00 |
Frozen Mouse Embryo | B6.129P2(C)-Arhgef11<tm1.1Mak>/J | $2595.00 |
Frozen Mouse Embryo | B6.129P2(C)-Arhgef11<tm1.1Mak>/J | $3373.50 |
Frozen Mouse Embryo | B6.129P2(C)-Arhgef11<tm1.1Mak>/J | $3373.50 |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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