STOCK Aire^{tm4.1(AIRE/GFP)Mmat}/J

Stock number: 036496
Product name: Aire/AGF-KI
Research areas: Immunology, Inflammation and Autoimmunity Research

Description

Aire/AGF-KI (also called Aire hAGF) is a knock-in/knock-out allele with the human AIRE gene, a green fluorescent protein (GFP), and a Flag tag inserted into exon 1 of the endogenous mouse Aire locus. These mice may be useful for identifying and tracing cell lineages of medullary thymic epithelial cells (mTECs) and in the study of immunology.

Detailed description

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

Aire/AGF-KI (also called Aire hAGF) is a knock-in/knock-out allele with the human AIRE gene, a green fluorescent protein (GFP), and a Flag tag inserted into exon 1 of the endogenous mouse Aire locus.

When crossed with Aire KO mice (see Stock No. 036465) to generate mice homozygous for the Aire KO allele and heterozygous for the hAGF knock-in/knock-out allele, the resulting mice fail to exhibit severe pneumonitis, a lethal autoimmune phenotype typical of Aire-deficient NOD mice, but do show autoimmune reactions against pancreatic acinar cells, and thus exhibit overall hypomorphic Aire function. Immunohistochemistry confirms GFP expression in the thymic medulla, with expression restricted to the cell nucleus in many medullary thymic epithelial cells (mTECs). Flow cytometry analysis reveals hAGF expression in ~65% of mTECs of heterozygotes, and in ~80% of mTECs in homozygotes (Kawano et al., 2015).

The donating investigator has made several Aire mouse lines available: Aire-KO (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/AGF-KI knock-in/knock-out allele (also called Aire-hAGF) 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 the human AIRE gene, along with a green fluorescent protein (GFP) reporter and Flag tag. The vector was electroporated into B6.Cg-Thy1^a-derived Bruce 4 embryonic stem (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-hAGF 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-hAGF mice. The resulting Aire-hAGF colony was then interbred prior to sending embryos to The Jackson Laboratory Repository. Upon arrival, the strain was recovered and maintained by breeding mutant mice to wildtype mice from the colony or to C57BL/6J mice (Stock No. 000664).

Allele

Symbol: Aire^{tm4.1(AIRE/GFP)Mmat}
Name: targeted mutation 4.1, Mitsuru Matsumoto
MGI accession ID: MGI:6771511
Generation method: Targeted
Attributes: Reporter; Null/Knockout; Inserted expressed sequence; Humanized sequence; Epitope tag
Synonyms: Aire/hAGF-KI; Aire^hAgf
Marker symbol: Aire
Marker name: autoimmune regulator (autoimmune polyendocrinopathy candidiasis ectodermal dystrophy)
Marker synonyms: AIRE1; APECED; APS1; APSI; PGA1
Chromosome: 10
Strain of origin: B6.Cg-Thy1^a
Molecular note: The targeting vector is designed to express a human AIRE /green fluorescent protein (GFP) reporter/Flag tag fusion protein followed by a neomycin selection cassette inserted between exons 1 and 2. Cre-mediated recombination removed the floxed neo cassette.