NOD.Cg-Aire^tm7Mmat/J

Stock number: 036558
Product name: 3xAire-KI
Research areas: Immunology, Inflammation and Autoimmunity Research

Description

3xAire-KI mice have 2 additional copies of the mouse Aire gene inserted into the endogenous Aire locus and dose-dependently over-express Aire protein. These mice may be useful in studying immunology and self-tolerance.

Detailed description

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

The 3xAire knock-in allele (also called 3xAire-KI) have 2 additional copies of the murine Aire gene, each preceded by a self-cleaving 2A peptide sequence (P2A), and inserted immediately after the final exon of the endogenous Aire gene. This is designed to have the endogenous Aire promoter/enhancer elements directing AIRE protein overexpression primarily in medullary thymic epithelial cells (mTECs).
The donating investigator reports 2x higher Aire mRNA levels in mTECs of heterozygotes and 3x higher in homozygotes, and similar dose-dependent increases in AIRE protein as determined by flow cytometry. 3xAire-KI mice exhibit normal development of CD4 and CD8 single-positive thymocytes, though 3xAire-KI showed a defect in negative selection and antigen-specific T-reg production using a TCR-transgenic model in which 3xAire-KI mice were crossed to OT-II mice [Stock No. 004194] (Nishijima et al., 2022).
The donating investigator reports that the presence of the 3xAire-KI allele prevents the development of the diabetic phenotype associated with the NOD genetic background.

Of note: The 3xAire-KI allele is also available on a congenic C57BL/6 genetic background as Stock No. 036693.

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 3xAire knock-in allele (also called 3xAire-KI) was created in the laboratory of Dr. Mitsuru Matsumoto (Institute for Enzyme Research, Tokushima University). The targeting vector, containing two sets of self-cleaving 2A peptide sequence followed by mouse Aire cDNA and a polyadenylation sequence, was targeted to the 3' end of the endogenous Aire locus on chromosome 10, after exon 14. Therefore, the construct reads Aire::P2A::Aire::P2A::Aire::PolyA. The construct 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/6JJcl mice for germline transmission. The resulting 3xAire-KI colony was then backcrossed to NOD/ShiJcl mice for at least 8 generations 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 NOD/ShiLtJ mice (Stock No. 001976).

Allele

Symbol: Aire^tm7Mmat
Name: targeted mutation 7, Mitsuru Matsumoto
MGI accession ID: MGI:6771325
Generation method: Targeted
Attributes: Inserted expressed sequence
Synonyms: 3xAire-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 contains two sets of self-cleaving 2A peptide sequence followed by mouse Aire cDNA and a polyadenylation sequence, was targeted to the 3' end of the gene (after exon 14).