NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Hprt1^b-m3/EshJ

Stock number: 012480
Product name: NSG hprt-null
Strain synonyms: NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Hprt^b-m3/EshJ
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Metabolism Research, Neurobiology Research, Research Tools, Virology Research

Description

These mutant mice combine the immunological characteristics of the NOD/ShiLtJ background, the severe combined immune deficiency mutation (scid), IL2 receptor gamma chain deficiency and a hypoxanthine phosphoribosyl transferase 1 (Hprt1^b-m3) null allele. This mutant mouse strain may be useful for xenotransplantation and drug screening.

Detailed description

Mice that are homozygous/hemizygous for these alleles are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. These mutant mice combine the following characteristics: NK cell deficiency (NOD/ShiLtJ background), T and B cell deficiency (Prkdc^scid), reduced numbers of lymphocytes and myeloid dendritic cells (Il2rg^tm1Wjl) and biochemically defective stromal cells (Hprt1^bm-3). These mice can be used for transplantation of human primary cancers. During tumor growth, Hprt1-defective murine fibroblast and stromal cells replace human stromal cells. In culture, the addition of HAT selection media eliminates murine fibroblasts and other stromal cells and facilitates the isolation of human cancer cells. These mice may be used to generate patient-specific low passage cell lines from primary cancers for use in chemosensitivity profiling and other applications.

Development

The Hprt1^bm-3 allele, a spontaneous 36 kb deletion at the 3' end of exon 2, was selected from an HPRT1 deficient 129P2/OlaHsd-derived E14TG2a embryonic stem cell line (ES). The X-linked mutation was introgressed into C57BL/6 mice for at least 5 generations. A heterozygous female was crossed to a NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ (Stock No. 005557) male; the backcross was continued for five generations. Correctly genotyped pups from each generation were selected using marker assisted genome scan analysis (speed congenic).

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: Hprt1^b-m3
Name: hypoxanthine guanine phosphoribosyl transferase B, mutation 3
MGI accession ID: MGI:1857299
Generation method: Spontaneous
Synonyms: Hprt^-
Marker symbol: Hprt1
Marker name: hypoxanthine phosphoribosyltransferase 1
Marker synonyms: HGPRT; Hgprtase; Hprt
Chromosome: X
Strain of origin: 129P2/OlaHsd
Molecular note: The allele contains a ~55 kb deletion spanning the promoter and first 2 exons. Subsequent direct sequence comparison with wild type DNA defined the exact breakpoints of the deletion, which lies 415 bp after the 3' end of exon 2, and determined the deletion size to be 36 kb (chrX:52,052,497-52,088,500 MM39).
General note: HPRT- embryonic stem cells were obtained by selecting for spontaneous mutation by incubation in medium containing 6-thioguanine. HPRT- males have no overt phenotype of abnormal behavior (J:15483). The mutation is due to a large deletion in the Hprt gene. In situ hybridization studies showed HPRT mRNA in high levels in most neurons, but not in glial cells, in normal mice. No HPRT mRNA was detected in the brains of male mice carrying this deletion (J:2058). Mutant mice have no HPRT detectable by Western blot analysis and no detectable HPRT enzyme activity in brain homogenates. They appear to have normal brain purine content, but de novo purine synthesis is accelerated four- to fivefold (J:11842). The Hprt^b-m3 mutation has been used in preimplantation studies to determine when the maternal and paternal alleles of Hprt are activated during early embryonic development (J:2389). Either administration of amphetamine (J:1847) or inhibition of adenine phosphoribosyltransferase (APRT) activity (J:4123) stimulates locomotor and stereotypic behaviors in HPRT-deficient mice. However, the null mutant for both Hprt and Aprt does not show the characteristics of Lesch-Nyhan disease (J:35822). Cells from mice hemizygous or homozygous for this mutation are HPRT-deficient and resistant to the drug 6-thioguanine (6TG).