STOCK Madd^tm1Bpra/J

Stock number: 030200
Product name: KMA1 Flox
Research areas: Apoptosis Research, Cell Biology Research, Diabetes and Obesity Research, Research Tools

Description

KMA1^flox/flox mice possess loxP sites flanking exons 3-4 of the solute carrier family 6 (neurotransmitter transporter, betaine/GABA), member 12 gene making them useful for studying glucose-induced insulin secretion.

Detailed description

KMA1^flox/flox mice have loxP sites flanking exons 3-4 of the MAP-kinase activating death domain (Madd) gene. MADD is a death domain-containing adaptor protein that interacts with the death domain of TNF-alpha receptor 1 to activate mitogen-activated protein kinase (MAPK) and transmit apoptotic signaling. MADD also plays a role in regulating cell proliferation, survival and death through alternative mRNA splicing. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exons 3-4 deleted in cre-expressing tissues.

When these mice are bred with Tg(Ins2-cre/ERT)1Dam mice (See Stock Nos. 008122 and 023972) expressing Cre Recombinase in pancreatic β-cells, the resulting offspring are hyperglycemic and glucose intolerant due to reduced and delayed glucose-induced insulin production.

Development

A targeting vector was designed to insert a single loxP site, followed by a frt-flanked neomycin resistance (neo) cassette, upstream of exon 3, and a second loxP site downstream of exon 4 of the MAP-kinase activating death domain (Madd) gene. The construct was electroporated into 129P2/OlaHsd-derived E14 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and resulting chimeric males were bred with C57BL/6 females. The neo cassette was removed by breed offspring to mice expressing Flp Recombinase on a C57Bl/6 genetic background. Resulting KMA1^flox mice were bred to C57BL/6 mice for at least 2 generations and the Flp-expressing transgene was selected away during the process. 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: Madd^tm1Bpra
Name: targeted mutation 1, Bellur S Prabhakar
MGI accession ID: MGI:5704429
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Marker symbol: Madd
Marker name: MAP-kinase activating death domain
Chromosome: 2
Strain of origin: 129P2/OlaHsd
Molecular note: An FRT-flanked neomycin resistance cassette with a 5' loxP site was inserted upstream of exon 3. An additional loxP site was inserted downstream of exon 4.