STOCK Aire^{tm5.1(HBEGF/GFP)Mmat}/J

Stock number: 036497
Product name: Aire/DTR-KI
Research areas: Immunology, Inflammation and Autoimmunity Research

Description

Aire/DTR/GFP knock-in/knock-out mice have the diptheria toxin receptor and green fluorescent protein (GFP) coding sequences 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 allow for the inducible ablation of Aire-expressing cells 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/DTR-KI knock-in/knock-out allele (also called Aire-DTR/GFP) has the diphtheria toxin receptor coding sequence and a green fluorescent protein (GFP) reporter in place of exon 1 of the Aire gene, resulting in disruption of Aire gene function and expression of GFP in Aire-expressing cells, as well as inducible cell ablation. Immunohistochemistry reveals GFP expression in the cytoplasm and nucleus of Aire-expressing medullary thymic epithelial cells (mTECs). A single administration of 500ng of diptheria toxin, or once daily administration for 5 consecutive days, results in dramatic loss of Aire⁺ mTECs and this cell population recovers approximately 3-4 days post treatment (Kawano et al., 2015).

The donating investigator 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/DTR-KI knock-in/knock-out allele (also called Aire-DTR/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 Diphtheria toxin receptor and green fluorescent protein, followed by a loxP-flanked neo cassette. 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-DTR/GFP 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-DTR/GFP mice. The resulting Aire-DTR/GFP colony was then interbred prior to sending embryos to The Jackson Laboratory Repository in 2021. 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^{tm5.1(HBEGF/GFP)Mmat}
Name: targeted mutation 5.1, Mitsuru Matsumoto
MGI accession ID: MGI:6771514
Generation method: Targeted
Attributes: Reporter; Null/Knockout; Inserted expressed sequence
Synonyms: (DTR)-KI; Aire/DTR-KI
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 Diptheria Toxin receptor (HBEGF)/ green fluorescent protein (GFP) reporter fusion protein followed by a neomycin selection cassette inserted between exons 1 and 2. Cre-recombination removed the floxed neomycin selection cassette.