B6(129S4)-Adra1a^{tm1c(EUCOMM)Hmgu}/DbeJ

Stock number: 041252
Product name: Adra1a^cKO
Research areas: Cardiovascular Research, Metabolism Research, Neurobiology Research, Research Tools

Description

Adra1a^cKO mice possess loxP sites flanking exon 3 of the Adra1a gene. These mice may be useful when studying cardiovascular function and stress response pathways.

Detailed description

Alpha-1-adrenergic receptors (coded by Adra1a) are members of the G protein-coupled receptor superfamily and are involved in regulating blood pressure, heart rate, and stress response pathways. These Adra1a conditional knock-out mice (Adra1a^cKO) have loxP sites flanking exon 3 of the Adra1a gene.

Heterozygous and homozygous mice are viable and fertile with no overt phenotype. When bred to mice that express tissue-specific Cre recombinase, the removal of exon 3 results in the possible truncation of the seventh transmembrane domain, the intracellular C-terminus, and 3 out of 17 key amino acid residues for the putative ligand binding site which may preserve partially functional receptors [PMID: 33268792].



These mice may be useful when studying adrenergic-regulation of cardiovascular and stress response pathways.

Of note, Adra1a^fl mice (Stock No. 041500) have loxP sites flanking the first coding exon (exon 2). When bred to mice that express tissue-specific Cre recombinase, Alpha-1A receptor protein is absent.

Development

C57BL/6N-derived JM8A3.N1 embryonic stem (ES) cells bearing the Adra1a^{tm1a(EUCOMM)Hmgu} targeted allele were obtained from The European Conditional Mouse Mutagenesis Program (EUCOMM). The targeting cassette is composed of an FRT site followed by lacZ sequence and an initial loxP site upstream of exon 3 of the adrenergic receptor, alpha 1a (Adra1a) gene. The first loxP site is followed by a neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of exon 3. Targeted ES cells were injected into recipient blastocysts. The resulting chimeric mice were bred with FLP-Recombinase mice (Stock No. 012930) to excise the FRT-flanked lacZ-loxP-neo cassette. Adra1a conditional knock-out (Adra1^cKO) mice, with neo cassette removed and floxed exon 3 intact, were maintained on a C57BL/6N background by the donating laboratory. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

Allele

Symbol: Adra1a^{tm1c(EUCOMM)Hmgu}
Name: targeted mutation 1c, Helmholtz Zentrum Muenchen GmbH
MGI accession ID: MGI:6502595
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: Adra1a^cKO
Marker symbol: Adra1a
Marker name: adrenergic receptor, alpha 1a
Marker synonyms: Adra1c; ADRA1L1; ALPHA1AAR
Chromosome: 14
Strain of origin: C57BL/6N-A^tm1Brd
Molecular note: The L1L2_Bact_P cassette was inserted at position 66964438 of Chromosome 14 upstream of the critical exon(s) (Build GRCm39). The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by a neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of the targeted exon(s) at position 66966027. The critical exon(s) is/are thus flanked by loxP sites. A "conditional ready" (floxed) allele was created by flp recombinase expression in mice carrying this allele to remove the lacZ sequence and neo selection cassette, leaving loxP sites flanking the critical exon(s). Further information on targeting strategies used for this and other IKMC alleles can be found at http://www.informatics.jax.org/mgihome/nomen/IKMC_schematics.shtml