NOD.Cg-Prkdc^scid Otc^em1Bshep Il2rg^tm1Wjl/J

Stock number: 034569
Product name: NSG.Otc^R129H
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Metabolism Research, Mouse/Human Gene Homologs, Research Tools, Virology Research

Description

NSG.Otc^R129H knock-in mice carry a R129H (arginine to histidine) substitution in the X-linked Otc (ornithine transcarbamylase) gene on an immunodeficient NSG background. This strain may be useful for studying early-onset ornithine transcarbamylase deficiency.

Detailed description

CRISPR/cas9 endonuclease-mediated genome editing of the X-linked Otc (ornithine transcarbamylase) gene was used to introduce an amino acid substitution at position 129 altering arginine to histidine (R129H). The R129H mutation has been identified in patients with early-onset ornithine transcarbamylase deficiency. The targeted Otc gene encodes a mitochondrial matrix enzyme. Missense, nonsense, and frameshift mutations in this enzyme lead to ornithine transcarbamylase deficiency, which causes hyperammonemia. Heterozygous Otc^R129H females on the immunodeficient NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ (NSG) background are viable and fertile. Hemizygous NSG-Otc^R129H males are viable, fertility has not been characterized. Limited testing on hemizygous males at the The Jackson Laboratory indicates that there are normal levels of ammonia (NH₃) in plasma.

Development

CRISPR/cas9 endonuclease-mediated genome editing was used to create an amino acid substitution at position 129 altering arginine to histidine (R129H; CG to CA) in exon 4 of the X-linked Otc (ornithine transcarbamylase) gene. This mutation was identified as a human early-onset ornithine transcarbamylase deficiency associated variant allele. Single guide RNA and Cas9 endonuclease were introduced into NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ-derived fertilized eggs by pronuclear injection. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ mice (Stock No. 005557, NSG) for germline transmission. The colony was maintained on the NSG background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize NSG oocytes.

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: Otc^em1Bshep
Name: endonuclease-mediated mutation 1, Ben Shepherd
MGI accession ID: MGI:6384615
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Synonyms: Otc^R129H
Marker symbol: Otc
Marker name: ornithine transcarbamylase
Marker synonyms: OCTD; OTC1; OTCase; OTCD
Chromosome: X
Strain of origin: NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ
Molecular note: CRISPR/cas9 endonuclease-mediated genome editing was used to create an amino acid substitution at position 129 altering arginine to histidine (R129H; CG to CA) in exon 4. This mutation was identified as a human early-onset ornithine transcarbamylase deficiency associated variant allele.

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.