NOD.Cg-Tg(HLA-DRA*0101,HLA-DRB1*0101)1Dmz Prkdc^scid H2-Ab1^b-tm1Doi Il2rg^tm1Wjl/SzJ

Stock number: 030331
Product name: NSG Ab^o DR1
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG Ab^o DR1 mice lack murine MHC class II and express mutant human leukocyte antigen (HLA)-DR1 on the immunodeficient NOD scid gamma (NSG) background. This strain may be useful in studies utilizing human hematopoietic stem cell-engraftment methods to develop alternative therapeutic techniques for immune dysfunctions.

Detailed description

NSG Ab^o DR1 mice are NOD.scid.Il2Rγc^null ("NSG") animals also harboring the H2-Ab1 knock-out allele (Ab^o) and the human leukocyte antigen DR1 transgene (HLA-DR1).

NSG Ab^o DR1 females homozygous for all three mutations and homozygous for the HLA-DR1 transgene, and males homozygous for scid, homozygous for Ab^o, homozygous for HLA-DR1 and hemizygous for the X-linked Il2Rγc^null may be collectively referred to as homozygous NSG Ab^o DR1 mice.

Homozygous NSG Ab^o DR1 express HLA-DR1 and lack murine MHC class II, on an NSG background (see Stock No. 005557 for NSG mouse description). Homozygous NSG Ab^o DR1 mice are viable and fertile. These mice may be useful in studies utilizing human hematopoietic stem cell-engraftment methods to develop alternative therapeutic techniques for immune dysfunctions.

Development

The HLA-DRA*0101,HLA-DRB1*0101 (HLA-DR1) transgene was created as follows: A transgenic construct containing the mouse MHC Class II H2-Ea and H2-Eb1 genes engineered to encode proteins in which the alpha1 domain (amino acids 1-85) and the beta1 domain (amino acids 1-96) were replaced by the corresponding amino acids of the human MHC Class II protein encoded by HLA-DRA1*0101 and HLA-DRB1*0101. Mice were generated by injecting the construct into fertilized (C57BL/6XSJL/J)F2 oocytes. Founder mice were backcrossed for three generations to B10.M-H2^f and then intercrossed to fix the H2^f locus. Mice were subsequently bred onto an NSG background (Stock No. 005557; NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ).

To generate a colony of NSG Ab^o DR1 mice (Stock No. 030331), HLA-DR1 transgenic mice on the NSG background were bred to NSG mice harboring the mouse MHC class II knock-out allele (Ab^o; see Stock No. 021885).

The H2-Ab1 knock-out allele in Stock No. 030331 was originally reported to be H2-Ab1^b-tm1Gru (created by Dr. Michael J. Grusby; while at Harvard) - harboring a forward-oriented neomycin resistance cassette that disrupts H2-Ab1 exon 2 (and made in ES cells with a deletion in the promoter region of H2-Ea). In 2018, quality control efforts at The Jackson Laboratory were completed that revealed Stock No. 030331 instead has the very similar H2-Ab1^b-tm1Doi knock-out allele (created by Drs. Christophe Benoist and Diane Mathis; while at INSERM) - harboring a reverse-oriented neomycin resistance cassette that disrupts H2-Ab1 exon 2 (and made in ES cells with a deletion in the promoter region of H2-Ea).

Of note, Stock No. 030331 may be segregating for H2^f and H2^b alleles.

 

Allele

Symbol: Tg(HLA-DRA*0101,HLA-DRB1*0101)1Dmz
Name: transgene insertion 1, Dennis M Zaller
MGI accession ID: MGI:3809457
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: DR1; DR1 Tg; HLA-DR1; Tg(HLA-DRB1*01)1Dmz
Marker symbol: Tg(HLA-DRA*0101,HLA-DRB1*0101)1Dmz
Marker name: transgene insertion 1, Dennis M Zaller
Marker synonyms: DR1; DR1 Tg; Tg(HLA-DRB1*01)1Dmz
Chromosome: 5
Strain of origin: (C57BL/6 x SJL/J)F2
Expressed genes: HLA-DRA,major histocompatibility complex, class II, DR alpha,human; HLA-DRB1,major histocompatibility complex, class II, DR beta 1,human
Site of expression: virtually all cells in peripheral blood that express MHC Class II

Molecular note: A chimeric MHC class II complex was created by fusing the human peptide binding domains of class II alpha and beta chains to the CD4 binding domains of murine class II alpha or beta chains. The was done by the construction of two separate transgene constructs. The alpha1 and beta1 domains isolated from human DRA1*0101 and DRB1*0101 replaced the corresponding alpha1 and beta1 domains of the H2-Ea and H2-Eb1 genes that make up mouse I-E^d. The native murine promoters were left intact. These transgene constructs were co-injected into fertilized oocytes and the founder mice with the highest expression of the chimeric MHC was chosen for further propagation. Flow cytometry analysis indicates that virtually all cells in peripheral blood that express endogenous MHC-II also express the chimeric MHC-II. The transgene construct was later shown (in J:126700) to have inserted into chromosome 5.

Allele

Symbol: Prkdc^scid
Name: severe combined immunodeficiency
MGI accession ID: MGI:1857113
Generation method: Spontaneous
Synonyms: scid
Marker symbol: Prkdc
Marker name: protein kinase, DNA activated, catalytic polypeptide
Marker synonyms: DNAPDcs; DNAPK; DNA-PK; DNAPKc; DNA-PKC; DNA-PKcs; DNPK1; DOXNPH; dxnph; HYRC; HYRC1; IMD26; p350; S473K; slip; XRCC7
Chromosome: 16
Strain of origin: C.BKa-Igh^b/Icr
Site of expression: T and B lymphocytes.
Molecular note: A T-to-A transversion point mutation at a position corresponding to codon 4046 (codon 4095 in transcript ENSMUST00000023352.8) created a premature stop codon (p.Y4046*).

Allele

Symbol: Il2rg^tm1Wjl
Name: targeted mutation 1, Warren J Leonard
MGI accession ID: MGI:1857455
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: [KO]gammac; CD132^-; gammac^-; gc^-; Il2rg^tm1Wjll; IL2Rgamma^null
Marker symbol: Il2rg
Marker name: interleukin 2 receptor, gamma chain
Marker synonyms: [g]c; Ab2-183; Cd132; CIDX; common cytokine receptor gamma chain; common gamma chain; gamma C receptor; gamma(c); gammaC; gc; IL-2RG; IMD4; P64; SCIDX; SCIDX1
Chromosome: X
Strain of origin: 129S4/SvJae
Site of expression: Primarily lymphoid cells.
Molecular note: A neomycin resistance cassette replaced part of exon 3 and all of exons 4 - 8 of the gene, resulting in the loss of most of the extracellular domain and all of the transmembrane and cytoplasmic domains of the protein.

Allele

Symbol: H2-Ab1^b-tm1Doi
Name: targeted mutation 1, Christophe Benoist and Diane Mathis
MGI accession ID: MGI:1927483
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: A beta⁰; A^bbeta^-; AB0; Abeta-; Class II⁰; H2-Ab1^tm1Dim; H2-Ab1^tm1Doi; H2-Ab1tm1Doi; II⁰; MHC class II-; MHCII^-; MHC-II-k.o.
Marker symbol: H2-Ab1
Marker name: histocompatibility 2, class II antigen A, beta 1
Marker synonyms: A beta; Abeta; Bb; CELIAC1; DQB2; H-2Ab; H2-Ab; HLA-DQB; HLA-DQB1; HLA-DXB; I-A^b; Ia2; Ia-2; IAb; I-Abeta; IDDM1; Rmcs1; RT1.B
Chromosome: 17
Strain of origin: 129S2/SvPas
Molecular note: The second exon was disrupted by the insertion of a neomycin resistance gene. In addition, the ES cell line used was derived from the 129S2/SvPas strain, which carries a deletion in the promoter region of H2-Ea. Consequently, these MHC class II molecule-deficient mice lacked cell surface expression of both class II-A and class II-E MHC proteins.