B6.Cg-Aire^tm2Mmat/EhrlJ

Stock number: 036498
Product name: Aire/GFP-KI
Strain synonyms: Aire^gfp, B6.Cg-Aire^tm2Mmat/J
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Aire/GFP-KI knock-in/knock-out mice have a green fluorescent protein (GFP) coding sequence inserted into exon 1 of the autoimmune regulator (autoimmune polyendocrinopathy candidiasis ectodermal dystrophy) (Aire) locus. These mice are designed to have the endogenous Aire promoter/enhancer sequences directing expression of enhanced green fluorescent protein primarily in medullary thymic epithelial cells (mTECs) as well as to disrupt expression of the Aire gene and may be useful in the study of autoimmunity.

Detailed description

The Aire gene encodes an E3 ubiquitin ligase which plays a critical role in immune self-tolerance induction.

The Aire/GFP-KI knock-in/knock-out allele (also called Aire^gfp) has green fluorescent protein (GFP) coding sequence in place of exon 1 of the Aire gene, resulting in disruption of Aire gene function and expression of GFP in Aire-expressing cells. Mice homozygous or heterozygous for the Aire^gfp allele express GFP in a pattern consistent with endogenous Aire expression, primarily in medullary thymic epithelial cells. Homozygous Aire^gfp mice exhibit altered cellular morphology in the thymus beginning as early as postnatal day 1 (Yano et al., 2008 J Exp Med 205:2827).

Dr. Mitsuru Matsumoto has made several Aire mouse lines available: Aire-KO (Stock No. 036465), Aire/AGF-KI (Stock No. 036496), Aire/DTR-KI (Stock No. 036497), Aire/GFP-KI (Stock No. 036498), Aire/OVA-KI (Stock No. 036500), NOD 3xAire-KI (Stock No. 036558), B6 3xAire-KI (Stock No. 036693), Aire/CreER-BAC Tg (Stock No. 036499) and hAIRE-Tg (Stock No. 036559).

Development

The Aire/GFP-KI knock-in/knock-out allele (also called Aire^gfp) was created in the laboratory of Dr. Mitsuru Matsumoto (Institute for Enzyme Research, Tokushima University). First, a targeting vector was designed to replace exon 1 of the autoimmune regulator (autoimmune polyendocrinopathy candidiasis ectodermal dystrophy) (Aire) locus on chromosome 10 with a green fluorescent protein (GFP) reporter and loxP-flanked neo cassette. The vector was electroporated into (C57BL/6 x CBA)F1-derived TT2 ES cells. Correctly targeted ES cells were injected into recipient blastocysts, and chimeric mice were bred with C57BL/6 mice for germline transmission. Offspring were bred with Ayu1-cre-expressing mice for germline deletion of the loxP-flanked neo cassette. The resulting Aire/GFP-KI colony was then crossed to C57BL/6 mice for removal of the Ayu1-cre allele, and the subsequent progeny were intercrossed to generate homozygous Aire^gfp mice. In 2013, mice were sent to Dr. Lauren I.R. Ehrlich (The University of Texas at Austin), where they were maintained by breeding to C57BL/6J as well as other C57BL/6 mutant strains. In 2024, Dr. Ehrlich sent Aire^gfp heterozygous mice (harboring no other mutant alleles) on a C57BL/6 genetic background to The Jackson Laboratory Repository. 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: Aire^tm2Mmat
Name: targeted mutation 2, Mitsuru Matsumoto
MGI accession ID: MGI:3818933
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Synonyms: Aire^gfp
Marker symbol: Aire
Marker name: autoimmune regulator (autoimmune polyendocrinopathy candidiasis ectodermal dystrophy)
Marker synonyms: AIRE1; APECED; APS1; APSI; PGA1
Chromosome: 10
Strain of origin: (C57BL/6NCrlj x CBA/JNCrlj)F1
Molecular note: Genomic sequence from immediately after the Kozak sequence in exon 1 to the exon 2 was replaced with GFP and a floxed neo cassette via homologous recombination. The neo cassette was removed via in vivo Cre mediated recombination. Immunohistochemical analysis in heterozygous mice indicates that the pattern of GFP expression is similar to the pattern of endogenous gene expression.