NOD.Cg-Rag1^tm1Mom Fah^em1Mvw Il2rg^tm1Wjl/MvwJ

Stock number: 018454
Product name: FAH^Δexon9 NRG
Strain synonyms: FNRG, FNRG
Research areas: Cancer Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools

Description

These FNRG mice contain a ZFN-mediated mutation that disrupts the Fah gene, and knock-out alleles of the Rag1 and Il2rg genes. This strain may be useful as a xenorecipient platform for human hepatocytes.

Detailed description

FAH^Δexon9 NRG mice contain a ZFN-mediated knock-out mutation targeted to exon 9 of the Fah fumarylacetoacetate hydrolase gene on the NRG background (NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/SzJ). The 172 base pair deletion mutation in exon 9 of Fah causes a premature stop. Heterozygotes are viable and fertile. Homozygotes develop a wasting phenotype unless fed a special diet (SSNIFF: S9541-E006) that lacks tyrosine and restricts phenylalanine (0.6%) and provided water containing nitisinone, 2-(2- nitro-4-trifluoro-methylbenzoyl)-1,3-cyclohexanedione, (NTBC) 16 mg/L. The special diet is a modified Baker Diet SSNIFF: S9541-E006, produced by SSNIFF (Germany). Histopathological examination of liver and kidney from homozygotes not treated with NTBC shows renal and hepatocyte cell and tissue injury. Successful human hepatocyte engraftment is detected by quantification of serum human albumin levels (ELISA) and immunohistochemistry. Fewer engrafted male FAH^Δexon9 NRG mice (homozygous for the Rag1 and Fah alleles and hemizygous for the Il2rg allele) survive compared to female FAH^Δexon9 NRG mice due to accumulation of toxic tyrosine catabolism metabolites.

Development

Zinc finger nuclease (ZFN) editing induced a 172 base pair deletion mutation in exon 9 of the Fah gene. The ZFN mRNAs were injected into NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/SzJ zygotes (Stock No. 007799). Founder line 1207, carrying a 172 base pair deletion mutation in exon 9, which generates a premature STOP codon, was selected for further study. Sperm was cryopreserved. To establish a live colony, an aliquot of frozen sperm was used to fertilize NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/SzJ oocytes (Stock No. 007799).

Allele

Symbol: Rag1^tm1Mom
Name: targeted mutation 1, Peter Mombaerts
MGI accession ID: MGI:1857241
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Rag1
Marker name: recombination activating gene 1
Chromosome: 2
Strain of origin: 129S7/SvEvBrd-Hprt⁺
Site of expression: expression is seen in bone marrow derived cell lines.
Molecular note: A 1356 bp genomic fragment of the Rag1 gene, encoding the nuclear localization signal and the zinc-finger motif, was replaced by a neomycin cassette. A mutant transcript expressed from this allele was detected by Northern blot in bone marrow derived cell lines from homozygous mice.

Allele

Symbol: Il2rg^tm1Wjl
Name: targeted mutation 1, Warren J Leonard
MGI accession ID: MGI:1857455
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Il2rg
Marker name: interleukin 2 receptor, gamma chain
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: Fah^em1Mvw
Name: endonuclease-mediated mutation 1, Michael Wiles
MGI accession ID: MGI:5485380
Generation method: Endonuclease-mediated
Attributes: Null/Knockout
Marker symbol: Fah
Marker name: fumarylacetoacetate hydrolase
Chromosome: 7
Strain of origin: NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/SzJ
Molecular note: Zinc finger nuclease (ZFN) editing induced a deletion of 172 bp in exon 9 of the Fah gene. The deletion results in a premature stop codon.