The Adam15 E>A knockin mice carry a point mutation that inactivates the catalytically active zinc-dependent metalloprotease domain resulting reduced tumor development in a heterotopic tumor injection model.
Carl P Blobel, Hospital for Special Surgery-Weill Med
Adam15 or a disintegrin and metallopeptidase domain 15 (metargidin) encodes a membrane-anchored metalloproteinase. Members of the ADAM family are involved in pathological neovascularization and are associated with cancer, atherosclerosis, RA and osteoarthritis. The Adam15 E>A knockin allele inactivates the catalytically active zinc-dependent metalloprotease domain. Mice homozygous for the mutation are viable and fertile. When injected heterotopically with melanoma cells, Adam15 E>A mice form fewer and smaller tumors than wild-type mice. This strain may be useful for studying the catalytic domain in Adam15 and tumor development.
The targeting vector was designed to insert an E>A point mutation into exon 11 by site-directed mutagenesis. An FRT-flanked neomycin cassette was inserted downstream of exon 11. The mutation results in an A to C change at nucleotide 6829 altering the corresponding amino acid from glutamate to alanine.
The construct was electroporated into unspecified embryonic stem (ES) cells derived from 129P2/OlaHsd mice. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to C57BL/6J females. Mice were crossed to B6;SJL-Tg(ACTFLPe)9205Dym/J to remove the neomycin cassette. Heterozygous mice were mated and offspring crossed to mixed C57BL/6 and 129/SvJ mice for several generations. Upon arrival, mice were bred to C57BL/6J for at least 1 generation to establish the colony.
|Allele Name||targeted mutation 2.1, Carl P Blobel|
|Allele Type||Targeted (Not Applicable)|
|Gene Symbol and Name||Adam15, a disintegrin and metallopeptidase domain 15 (metargidin)|
|Strain of Origin||129P2/OlaHsd|
|Molecular Note||The targeting vector is designed to insert a E to A point mutation into the catalytic domain (exon 11) by site-directed mutagenesis. An FRT-flanked neomycin cassette is inserted downstream of exon 11. The mutation results in a A to C change at nucleotide 6829 altering the corresponding amino acid from glutamate to alanine. Flp-mediated recombination removed the FRT-flanked neo cassette.|
While maintaining a live colony, these mice are bred as homoygotes.
When using the Adam15 E>A mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #37639 in your Materials and Methods section.