NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(CSF1)3Sz/J

Stock number: 028657
Product name: NSG-Quad; NSG-Q

Strain synonyms: NSG-SGM3 CSF1, NSG-Quad
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG QUAD mice express the human cytokines CSF1 [colony stimulating factor 1 (macrophage)], interleukin-3 (IL3; IL-3), granulocyte/macrophage-stimulating factor(CSF2; GM-CSF), and Steel factor (KITLG; SCF or SF) on the immunodeficient NSG background. In humanized NSG QUAD mice, the human cytokines can be used to support the survival of human induced pluripotent stem cells (hiPSCs).

Detailed description

NSG QUAD (NSG-Q) mice express human CSF1 [colony stimulating factor 1 (macrophage)] and the triple transgenic SGM3 coinjected transgenes, each with a human cytomegalovirus promoter/enhancer sequence driving expression of the human interleukin-3 gene (IL3; IL-3), the human granulocyte/macrophage-stimulating factor gene (CSF2; GM-CSF), and the human Steel factor gene (KITLG; SCF or SF) on the immunodeficient NSG (Stock No. 05557) background. NSG mice carry the Prkdc^scid and Il2rg^tm1Wjl alleles on the NOD/LtShiJ background; they should have no mature T cells or B cells, lack functional natural killer (NK) cells, have reduced numbers of lymphocytes and myeloid dendritic cells, and be deficient in cytokine signaling. These mice do not spontaneously develop diabetes. Mice that are homozygous for scid, hemizygous/homozygous for the X-linked Il2rg, homozygous for SGM3 and hemizygous for CSF1 are viable and fertile. Males homozygous for the CSF1 transgene do not produce viable sperm.

The CSF1 and SGM3 transgenes encode the human cytokines CSF1, IL3, GM-CSF (CSF2) and SCF, which function to support the stable engraftment of human myeloid lineages and regulatory T cell populations. SGM3 mice constitutively produce 2-4 ng/ml serum levels of human IL-3, GM-CSF, and SF. The NSG QUAD strain is an immunodeficient mouse model producing human cytokines that can be used to support the survival of human induced pluripotent stem cells (hiPSCs). Brains from NSG-QUAD mice transplanted with hiPSC-derived microglial precursors (iMPS) generate cells that develop microglial morphology and gene expression patterns similar to primary human microglia. In some brain areas, the donor cells represent up to 50% of the total number of microglia.

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Similar to other immunodeficient strains, maintaining NSG-QUAD mice in high health status (specific pathogen-free) vivaria promotes overall colony health. If NSG-QUAD animals are maintained in low health barrier rooms, the use of medicated water (e.g., sulfatrim/trimethoprim-sulfa or enrofloxacin/Baytril) is suggested to increase overall colony health.

Development

The CSF1 transgenic construct containing the human colony stimulating factor 1 (macrophage), CSF1, gene from the 157 Kb BAC clone RP11-19F3 was microinjected into the pronuclei of fertilized NOD.CB17-Prkdc^scid/J eggs. Founder line 3 was established and mated to NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/J (Stock No. 005557) mice.

For the SGM3 transgene, three separate transgenes were designed each carrying either the human interleukin-3 gene (IL3; IL-3), the human granulocyte/macrophage-stimulating factor gene (CSF2; GM-CSF), or the human Steel factor gene (KITLG; SCF or SF). Expression of each gene is driven by a human cytomegalovirus promoter/enhancer sequence, and is followed by a human growth hormone cassette and a polyadenylation (polyA) sequence. The transgenes were microinjected into fertilized C57BL/6xC3H/HeN oocytes. The resulting founders, carrying all three transgenes (3GS) were backcrossed to BALB/c-scid/scid mice for several generations and subsequently backcrossed to NOD.CB17-Prkdc^scid mice for at least 11 generations.

This strain was created by mating NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CSF1)3Sz/J (Stock No. 028654) mice and NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J (Stock No. 013062) mice. Progeny were interbred until all offspring were homozygous for scid, hemizygous/homozygous for the X-linked Il2rg targeted mutation and homozygous for the SGM3 and hemizygous for the CSF1 transgenes.

Allele

Symbol: Tg(CMV-IL3,CSF2,KITLG)1Eav
Name: transgene insertion 1, CJ Eaves
MGI accession ID: MGI:3653745
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: 3/GM/SF; SGM3; Tg(hSCF/hGM-CSF/hIL3
Marker symbol: Tg(CMV-IL3,CSF2,KITLG)1Eav
Marker name: transgene insertion 1, CJ Eaves
Chromosome: UN
Strain of origin: (C57BL/6 x C3H/HeN)F1
Expressed genes: IL3,interleukin 3,human; CSF2,colony stimulating factor 2,human; KITLG,KIT ligand,human
Molecular note: cDNAs encoding human interleukin 3, colony stimulating factor 2 (granulocyte-macrophage) and kit ligand were independently ligated into expression cassettes in which the cDNA resides downstream of the cytomegalovirus (CMV) promoter/enhancer sequence and is followed by human growth factor intronic and polyadenylation sequences. A transgenic mouse that was shown by ELISA to have serum levels of all three growth factors at ng/ml levels was used to establish a line with the same phenotype.

Allele

Symbol: Prkdc^scid
Name: severe combined immunodeficiency
MGI accession ID: MGI:1857113
Generation method: Spontaneous
Synonyms: SCID; scid
Marker symbol: Prkdc
Marker name: protein kinase, DNA activated, catalytic polypeptide
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: gammac^-; IL2Rgamma^null; [KO]gammac; CD132^-; gc^-; Il2rg^tm1Wjll
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: Tg(CSF1)3Sz
Name: transgene insertion 3, Leonard Shultz
MGI accession ID: MGI:5699722
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(CSF1)3Sz
Marker name: transgene insertion 3, Leonard Shultz
Chromosome: UN
Strain of origin: NOD.Cg-Prkdc^scid
Expressed genes: CSF1,colony stimulating factor 1,human
Site of expression: CSF1 encodes a cytokine involved in the regulation of macrophages.
Molecular note: This transgene consists of a 157 kb human BAC clone, RP11-19F3, containing the entire genomic sequence of the human CSF1 gene including the endogenous promoter. CSF1 is the only gene in the plasmid.