NOD.Cg-Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav Tg(HLA-A/H2-D/B2M)1Dvs/J

Stock number: 035844
Product name: NSGF-SGM3‐A2 , SGM3F-A2
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSGF-SGM3‐A2 (also called SGM3F-A2) is an enhanced NOD scid gamma (NSG™) mouse model. In addition to features of B and T cell deficiency (contributed by the Prkdc^scid mutation) and NK cell deficiency (associated with the Il2rg knock-out allele), these animals also carry a CRISPR/cas9-generated knock-out of the mouse Flt3 (FMS-like tyrosine kinase 3) gene, improving the engraftment of diverse hematopoietic lineages, and a human IL3 (interleukin 3)/GM‐CSF (CSF2, colony stimulating factor 2 (granulocyte-macrophage)/SCF (KITL, kit ligand) transgene driven by a CMV promoter to support the stable engraftment of myeloid lineages and regulatory T cell populations (SGM3). Additionally, these mice carry a transgene (HLA‐A2/HHD) expressing human HLA class 1 heavy and light chains (Tg(HLA-A2/H2-D/B2m)1Dvs/SzJ) that permits HLA-restricted human immune responses.

These NSGF‐SGM3‐A2 mice enable improved engraftment of HLA‐A2 matched hematopoietic lineages and are useful for immune‐oncology, immunology and infectious disease studies.

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Detailed description

NSGF-SGM3‐A2 (also called SGM3F-A2) is an enhanced NOD scid gamma (NSG™) mouse model. In addition to features of B and T cell deficiency (contributed by the Prkdc^scid mutation) and NK cell deficiency (associated with the Il2rg knock-out allele), these animals also carry a CRISPR/cas9-generated knock-out of the mouse Flt3 (FMS-like tyrosine kinase 3) gene, improving the engraftment of diverse hematopoietic lineages, and a human IL3 (interleukin 3)/GM‐CSF (CSF2, colony stimulating factor 2 (granulocyte-macrophage)/SCF (KITL, kit ligand) transgene driven by a CMV promoter to support the stable engraftment of myeloid lineages and regulatory T cell populations (SGM3). Additionally, these mice carry a transgene (HLA‐A2/HHD) expressing human HLA class 1 heavy and light chains (Tg(HLA-A2/H2-D/B2m)1Dvs) that permits HLA-restricted human immune responses. These mice permit improved engraftment of diverse hematopoietic lineages and are useful for immune‐oncology, immunology and infectious disease studies.

Flt3 encodes a receptor that is critical for development of the dendritic cells and monocytic lineages. This knock-out allele incorporates a 436 bp deletion between introns 2 and 3 which leads to exon 3 skipping and the production of a non‐functional mRNA.  The mutation enables an increase in the availability of FLT3L (Flt3 ligand) to human cells, thereby improving engraftment of diverse hematopoietic lineages.

These NSGF‐SGM3‐A2 mice enable improved engraftment of HLA‐A2 matched hematopoietic lineages and are useful for immune‐oncology, immunology and infectious disease studies.

SPF (specific pathogen-free) conditions are recommended. Please see our NSG housing and breeding information for additional details.

View our Resources for the NSG™ mouse model, including:

• NSG™ Mouse Model Portfolio
• Housing and Breeding Considerations for NSG™ Mice
• Frequently Asked NSG™ Questions
• Breakthrough Research Using NSG™
• Immunodeficient Mouse and Xenograft Host Comparisons
• Flow Cytometry Characterization Data for Immunodeficient JAX Strains

Development

NSG‐HLA‐A2/HHD mice (Stock No. 014570) were bred to NSGF‐SGM3 mice (Stock No. 035843) for 2 generations to establish this line of NSGF‐SGM3‐A2 mice (Stock No. 035844) .

NSG‐HLA‐A2/HHD mice (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(HLA-A/H2-D/B2M)1Dvs/SzJ, Stock No. 014570) mice carry 3 mutations: Prkdc^scid, X-linked Il2rg^tm1Wjl, and transgene Tg(HLA-A2/H2-D/B2m)1Dvs/SzJ, expressing human HLA class 1 heavy and light chains. The strain was generated by crossing NOD/ShiLtDvs-Tg(HLA-A/H2-D/B2M)1Dvs/J (Stock No. 006604) mice with NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ (Stock No. 005557) animals. The mice were crossed to NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ (Stock No. 005557) for 9 generations.

NSGF‐SGM3 mice (Stock No. 035843) were generated through a cross of NSG‐SGM3 mice (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ, Stock No. 013062) with NSGF mice (NOD.Cg‐Prkdc^scid Il2rg^tm1Wjl-Flt3^em1Akp/J, Stock No. 035842) for 2 generations.

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: Tg(HLA-A/H2-D/B2M)1Dvs
Name: transgene insertion 1, David V Serreze
MGI accession ID: MGI:3711057
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: HDD; HHD
Marker symbol: Tg(HLA-A/H2-D/B2M)1Dvs
Marker name: transgene insertion 1, David Serreze
Marker synonyms: HDD; HHD
Chromosome: UN
Strain of origin: NOD/ShiLtDvs
Expressed genes: B2M,beta-2-microglobulin,human; HLA-A,major histocompatibility complex, class I, A,human
Molecular note: This transgene encodes a human B2-microglubulin (B2M) covalently linked to the MHC class 1, alpha1 and alpha2 binding domains of the human HLA-A2.1 gene, and the alpha3, cytoplasmic and transmembrane domains of the murine H2-D^b. The construct was injected into fertilized NOD/ShiLtDvs oocytes. The construct was originally described in J:41077 and used to create a different transgenic line.
General note: The HDD construct was created and used to generate a differnet transgenic mouse line in J:41077.

Allele

Symbol: Flt3^em1Akp
Name: endonuclease-mediated mutation 1, Karolina Palucka
MGI accession ID: MGI:6470929
Generation method: Endonuclease-mediated
Attributes: Null/Knockout
Marker symbol: Flt3
Marker name: FMS-like tyrosine kinase 3
Marker synonyms: CD135; FLK2; FLK-2; Flt-3; STK1; wmfl
Chromosome: 5
Strain of origin: NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ
Molecular note: CRISPR/cas9 genome editing is used to delete a 436 bp region that incorporate exon 3 and leads to exon 3 skipping and the production of a non‐functional mRNA.

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.