NOD.Cg-Alox15^tm1.1Nadl/MirmJ

Stock number: 037552
Product name: NOD-12/15-LO^loxP, NOD:Alox15^loxP
Research areas: Diabetes and Obesity Research, Research Tools

Description

NOD:Alox15^loxP mice possess loxP sites flanking exons 2-5 of the arachidonate 15-lipoxygenase (Alox15) gene on the diabetes susceptible NOD/ShiLtJ background. These floxed mice are useful for tissue specific deletion of Alox15 when studying obesity and diabetes.

12/15-LO^loxP/loxP is available on the C57BL/6 background as Stock No. 031835.

Detailed description

12/15-lipoxygenase (12/15-LO, Alox15) is an enzyme involved in promoting adipocyte dysfunction. 12/15-LO oxygenates polyunsaturated fatty acids to generate proinflammatory lipid mediators such as eicosanoids, hepoxilins, lipoxins, and other molecules. NOD.Alox15^loxP mice possess loxP sites flanking exons 2-5 of the arachidonate 15-lipoxygenase (Alox15) gene. Mice that are homozygous for this allele are viable and fertile and develop spontaneous autoimmune diabetes in a similar timeframe to standard NOD mice. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exons 2-5 deleted in the cre-expressing tissues.

When NOD.Alox15^loxP mice are bred to mice expressing a myeloid cell specific Cre recombinase (NOD.129P2(B6)-Lyz2^tm1(cre)Ifo/NadlJ, Stock No. 019096), resulting offspring (NOD.Alox15^Δmyel) exhibit a significant reduction of Alox15 in peritoneal cells, but not in spleen and pancreatic islets. NOD.Alox15^Δmyel mice develop less diabetes than controls (12.5% of females and 0% of males by 25 weeks of age) with increased β mass, decreased insulitis, and reduced numbers of both proinflammatory macrophages and dendritic cells expressing IL1B.

12/15-LO^loxP/loxP is available on the C57BL/6 background as Stock No. 031835.

 

Development

The 12/15-LO^loxP targeting vector was designed by Dr. Jerry Nadler (Eastern Virginia Medical School) to insert loxP sites flanking exons 2-5 of the arachidonate 15-lipoxygenase (Alox15) gene, and a FRT-flanked neomycin selection cassette was inserted downstream of exon 5. The construct was electroporated into B6.Cg-Thy1^a-derived Bruce4 embryonic stem (ES) cells. Resulting ES cells were injected into blastocysts and resulting 12/15-LO^loxP mice were bred to a FLP deleter strain on a C57BL/6J genetic background to remove the neo cassette. Progeny were crossed to remove the Flp-expressing transgene, and offspring were bred to C57BL/6J mice for at least 10 generations. Mice were subsequently backcrossed onto the NOD/ShiLtJ (Stock No. 001976) background by Dr. Raghavendra Mirmira (University of Chicago) using the speed congenic breeding technique. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize NOD/ShiLtJ oocytes (Stock No. 001976).

Allele

Symbol: Alox15^tm1.1Nadl
Name: targeted mutation 1.1, Jerry Nadler
MGI accession ID: MGI:4889210
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Alox15
Marker name: arachidonate 15-lipoxygenase
Chromosome: 11
Strain of origin: B6.Cg-Thy1^a
Molecular note: Development of this allele was contracted to Ozgene in Australia. Exons 2 to 5 were flanked by loxP sites. An FRT flanked neomycin selection cassette has been removed.