NOD.Cg-Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J

Stock number: 035843
Product name: NSGF-SGM3, NSG-SGM3-Flt3KO, SGM3F
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSGF-SGM3 (also called SGM3F or NSG-SGM3-Flt3KO) 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). These mice permit improved engraftment of diverse hematopoietic lineages and are useful for immune‐oncology, immunology and infectious disease studies.

These NSGF-SGM3 mice yield a better engraftment of human hematopoietic progenitor cells (HPCs) in the blood than NSGF mice (Stock No. 035842) that lack the Tg(CMV‐IL3,CSF2,KITLG)1Eav allele. They also allow efficient engraftment of adult human HPCs. Moreover, they enable the generation of specific antibody responses to vaccination.

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Detailed description

NSGF-SGM3 (also called SGM3F or NSG-SGM3-Flt3KO) 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).

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 mice yield a better engraftment of human hematopoietic progenitor cells (HPCs) in the blood than NSGF mice (Stock No. 035842) that lack the Tg(CMV‐IL3,CSF2,KITLG)1Eav allele. They also allow efficient engraftment of adult human HPCs. Moreover, they enable the generation of specific antibody responses to vaccination.

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‐SGM3 mice (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ, Stock No. 013062) were bred to NSGF mice (Stock No. 035842) for 2 generations to establish the donating laboratory's colony of these NSGF-SGM3 (SGM3F, NSG-SGM3-Flt3KO, Stock No. 035843) mice.

NSGF mice (Stock No. 035842) were generated using the CRISPR/cas9 system. Cas9 and sgRNAs were co-injected into fertilized NOD.Cg‐Prkdc^scid Il2rg^tm1Wjl (Stock No. 005557) oocytes to delete a 436bp region incorporating exon 3 of the mouse Flt3 gene. The resulting founders were bred to NOD.Cg‐Prkdc^scid Il2rg^tm1Wjl mice, and then intercrossed until all offspring were homozygous for the Flt3 targeted mutation.

NSG‐SGM3 mice (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ, Stock No. 013062) were generated using three separate transgenes carrying either the human interleukin-3 gene (IL3), 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/6 x C3H/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 3GS and the 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. Thereafter, females homozygous for Prkdc^scid, homozygous for Il2rg^tm1Wjl, and homozygous for the 3GS transgenes were bred with males homozygous for Prkdc^scid, hemizygous for the X-linked Il2rg^tm1Wjl, and homozygous for the 3GS 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
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: 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.