NOD.Cg-Prkdc^scid H2-K1^b-tm1Bpe H2-Ab1^g7-em1Mvw H2-D1^b-tm1Bpe Il2rg^tm1Wjl/SzJ

Stock number: 025216
Product name: NSG-MHC I/II DKO
Strain synonyms: NSG-(K^bD^b)^null (IA)^null, NSG-(K^b D^b)^null (IA^null)
Research areas: Cancer Research, Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG-MHC I/II DKO mutant mice combine the features of the severe combined immune deficiency mutation (scid), IL2 receptor gamma chain deficiency, MHC class I molecule deficiency (H2-K and D), and MHC class II molecule deficiency (IA) and exhibit a significant delay in the onset of graft versus host disease (GVHD). They provide a model useful to study in vivo mechanisms of xenogeneic GVHD and to rapidly assess therapeutic agents.

These NSG-MHC I/II DKO mice are also called NSG-(K^bD^b)^null (IA)^null or NSG-(K^b D^b)^null (IA^null) in Brehm et al. 2019 FASEB J [PMID:30383447].

You can order this model engrafted with human PBMCs through our humanized mouse order form.

Detailed description

These NSG-MHC I/II DKO mice are also called NSG-(K^bD^b)^null (IA)^null or NSG-(K^b D^b)^null (IA^null) in Brehm et al. 2019 FASEB J [PMID:30383447]. These mice are NOD.scid;IL2rg-deficient (NSG) animals also containing null targeted mutations for mouse MHC class I genes (H2-K1 and H2-D1) and a TALEN-made null mutation for the mouse class II gene (H2-Ab1 [I-A]) - making them an MHC I/II double knock-out.

These NSG-MHC I/II DKO mice combine the features of the severe combined immune deficiency mutation or scid (B and T cell deficiency), IL2 receptor gamma chain deficiency (NK cell deficiency), MHC class I molecule deficiency (H2-K and D), and MHC class II molecule deficiency (IA). Mice homozygous for the five mutations are viable, normal in size and do not display any gross physical or behavioral abnormalities.

These mice exhibit a significantly delayed onset of xenogeneic GvHD compared to NSG mice. The rate and severity of onset is highly dependent on preparatory irradiation dose (or absence), specific donor used, and cell dose injected. Each PBMC donor requires a pilot engraftment using more than one cell dose to determine the overall engraftment characteristics. Once established, these engraftment characteristics are highly reproducible. At JAX, the survival of non-irradiated mice injected with 10 x 10⁶ human peripheral blood mononuclear cells (PBMC) is greater than 100 days, with 13/15 of mice exhibiting no symptoms of GVHD for up to 125 days. Because this strain carries a null mutation in the MHC class I H2-Ab1 gene rather than β-2M, IgG clearance is similar to that observed in the NSG model. This strain may be useful to study human immunity in the absence of GVHD and to evaluate antibody-based therapeutics.

Development

A targeting scheme was designed using a TALEN-mediated Non-Homologous End Joining (NHEJ) repair pathway to generate a null allele by targeting exon 2 of the MHC class II IAβ allele, H2-Ab1. TALEN mRNAs were introduced into zygotes from NOD.Cg-Prkdc^scid H2-K1^b-tm1Bpe H2-D1^b-tm1Bpe Il2rg^tm1Wjl/SzJ (Stock No. 023848). Progeny were intercrossed to generate females that are homozygous for the scid, H2-Ab1 (MHC class II) KO, H2-K1 KO, H2-D1 KO and Il2rg KO alleles. Males are homozygous for the scid, H2-Ab1 (MHC class II) KO, H2-K1 KO, H2-D1 KO and hemizygous for the X-linked Il2rg KO allele. All three H-2 alleles are closely linked on chromosome 17 and are likely to segregate together.

Allele

Symbol: H2-Ab1^g7-em1Mvw
Name: endonuclease-mediated mutation 1, Michael Wiles
MGI accession ID: MGI:6156109
Generation method: Endonuclease-mediated
Attributes: Null/Knockout
Synonyms: H2-Ab1^em1Mvw; IA^null
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: NOD.Cg-Prkdc^scid H2-K1^b-tm1Bpe H2-D1^b-tm1Bpe Il2rg^tm1Wjl/Sz
Molecular note: This TALEN-mediated null mutation has a 61 bp deletion in exon 2, Chr 17: :34264928-34264988 (GRCm38), followed by 39 bp of retained endogenous sequence, then a 507 bp deletion, Chr 17: 34265028-34265534, with a 7 bp insertion (CCGTCAC) in its place, the impact of which is a disruption of most of exon 2. Flow cytometric analysis confirmed this to be a null allele.

Allele

Symbol: H2-K1^b-tm1Bpe
Name: targeted mutation 1, Beatrice Perarnau
MGI accession ID: MGI:1931444
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: H2-K^b-; H-2K^o; H2-K1^tm1Bpe; K^-; K^b-
Marker symbol: H2-K1
Marker name: histocompatibility 2, K1, K region
Marker synonyms: AS; B-4901; CDA12; D6S204; H-2K; HLA-5.4; HLA-6.2; HLAA; HLAB; HLAC; HLA-CDA12; HLAF; HLA-JY3; HLC-C; MHC; MHC-G; PSORS1; QA1; RT1.A1(N); RT1.A2(N); RT1A-1; RT1-A1n; RT1A-2; RT1-A2n; RT1-C/E1; RT1-C/E3; RT1-C/E4; RT1-C/E5; RT1-C/E7; RT1-CE12; RT1-CE7; RT1-E2; rt1-El
Chromosome: 17
Strain of origin: 129P2/OlaHsd
Molecular note: The central part of the H2-K gene, including the second and third exons was replaced by a HPRT minigene.

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; 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-D1^b-tm1Bpe
Name: targeted mutation 1, Beatrice Perarnau
MGI accession ID: MGI:1931445
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: D^-; D^b-; H-2 D^bo; H-2D^b-; H-2D^b knockout; H-2D^o; H2-D1^tm1Bpe; HHD
Marker symbol: H2-D1
Marker name: histocompatibility 2, D region locus 1
Marker synonyms: AS; B-4901; CDA12; D6S204; H-2D; HLA-5.4; HLA-6.2; HLAA; HLAB; HLAC; HLA-CDA12; HLAF; HLAHP; HLA-JY3; HLC-C; MHC; MHC-G; PSORS1; QA1; RT1.A1(N); RT1.A2(N); RT1A-1; RT1-A1n; RT1A-2; RT1-A2n; RT1-C/E1; RT1-C/E3; RT1-C/E4; RT1-C/E5; RT1-C/E7; RT1-CE12; RT1-CE7; RT1-E2; rt1-El
Chromosome: 17
Strain of origin: 129P2/OlaHsd
Molecular note: Exons 1, 2 and 3 of the b haplotype gene were replaced with plasmid sequences and a neomycin cassette.