NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(IL15)1Sz/J

Stock number: 033216
Product name: NSG-SGM3-IL15
Strain synonyms: S15
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG-SGM3-IL15 mice expressing human IL3, GM-CSF (CSF2), SCF (KITLG), and IL15 combine the features of the highly immunodeficient NOD scid gamma (NSG) mouse with cytokines that support the stable engraftment of myeloid lineages, regulatory T cell populations and NK cells. As a result, these NSG-SGM3-IL15 mice allow superior engraftment of diverse hematopoietic lineages than other models and are useful for immuno-oncology, immunology and infectious disease studies.

CD34+ humanized NSG-SGM3-IL15 (Stock No. 703321) mice can be ordered on our humanized mouse order form or learn more about humanized NSG-SGM3-IL15 mice and see representative engraftment data here.

Detailed description

The NSG-SGM3-IL15 mice are NOD.scid.Il2Rγc^null ("NSG") animals with the 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), or the human Steel factor gene (KITLG; SCF or SF) and a human IL15 transgene.

NSG-SGM3-IL15 mice combine the features of the highly immunodeficient NSG (B, T, and NK deficiency) mouse with human cytokines (IL3, GM-CSF, SCF, and IL15) that support the stable engraftment of myeloid lineages, regulatory T cell populations and NK cells. Mice irradiated and injected with CD34+ human stems cells (HSC) develop significantly higher levels of functional human CD56+ NK cells in peripheral blood, spleen and bone marrow than NSG controls. Expression of SGM3 produces 2-4 ng/ml serum levels of human IL-3, GM-CSF, and SF. Mice homozygous for the four mutations are viable, normal in size and do not display any gross physical or behavioral abnormalities; survival is similar to NSG mice.

NSG-SGM3-IL15 mice allow superior engraftment of diverse hematopoietic lineages than other models and are useful for immuno-oncology, immunology and infectious disease studies.

Similar to other immunodeficient strains, maintaining NSG-SGM3-IL15 mice in high health status (specific pathogen-free) vivaria promotes overall colony health. If homozygous NSG-SGM3-IL15 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.

View our Resources for the NSG mouse model, including discussion forum, immunodeficient model comparison, flow cytometry data and categorized, up-to-date references.

Development

For the SGM3 allele, 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. These mice were bred to NOD.Cg-Prkdc^scid Il2rg^tm1Wjl mice and were then interbred until all offspring were homozygous for SGM3, scid and hemizygous/homozygous for the X-linked IL2rg targeted mutation. Upon arrival at The Jackson Laboratory, transgenic mice were bred to NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ mice (Stock No. 005557) for one generation to establish the colony. This colony is distributed as Stock No. 013062.

Mice were then bred with NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(IL15)1Sz/SzJ (Stock No. 030890) to create this strain. The IL15 transgenic construct consists of a 200 kbp human BAC RP11-620F3 containing the complete human IL15 (interleukin 15) gene.

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
Marker synonyms: 3/GM/SF
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
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: Tg(IL15)1Sz
Name: transgene insertion 1, Leonard Shultz
MGI accession ID: MGI:6201746
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: Tg(Hu-IL15)
Marker symbol: Tg(IL15)1Sz
Marker name: transgene insertion 1, Leonard Shultz
Chromosome: UN
Strain of origin: NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ
Site of expression: IL15 has widespread expression in a large number of cells and tissues.

Molecular note: The transgenic construct consists of a 200 kbp human BAC RP11-620F30 containing the complete human IL15 (interleukin 15) gene. Mice express a physiological level of human IL15 (7.1 +/- 0.3 pg/ml).