NOD.Cg-Il6^em1(IL6)Akp Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl/AkpJ

Stock number: 039496
Product name: NSGF6 , NSG-Flt3-KO-IL6-KI , NSGF-IL6ki
Strain synonyms: NSGF6
Research areas: Cancer Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG-Flt3-KO-IL6-KI (NSGF6) mice combine the features of the highly immunodeficient NOD scid gamma (NSG™) mouse (Stock No. 005557) with an FMS-like tyrosine kinase 3 (Flt3) knock-out and human interleukin 6 (IL6) knock-in/knock-out that support the stable engraftment of human hematopoiesis. These mice allow superior engraftment of diverse hematopoietic lineages and are useful for immuno-oncology, immunology, and infectious disease studies.

Please click here to order our Humanized Mice.

Detailed description

NSG-Flt3-KO-IL6-KI (NSGF6) immunodeficient mice, based on NOD scid gamma (NSG™) mice (Stock No. 005557), were developed to support long-term engraftment with human adult hematopoietic stem and progenitor cells (HSPCs). As both FMS-like tyrosine kinase 3 ligand (Flt3l) and interleukin 6 (IL6) are critical for many aspects of hematopoiesis, a knock-out of the mouse FMS-like tyrosine kinase 3 (Flt3) enzyme and a knock-in/knock-out of the human IL6 gene were incorporated. The resulting mice show an increase in the availability of mouse FLT3L to human cells and a dose-dependent production of human IL6 upon activation. Upon transplantation with a low number of human HSPCs from adult bone marrow, these humanized mice demonstrate a significantly higher engraftment with multilineage differentiation of human lymphoid and myeloid cells, and tissue colonization at one year after adult HSPC transplant. Female mice were used in the primary study for their anticipated capacity to better support human HSPC engraftment than males (see PMID:20207983 and PMID:26595075). Engraftment by males was not tested by the donating laboratory. These mice enable studies of human hematopoiesis and tissue colonization over time and may facilitate building autologous models for immuno-oncology 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-Flt3-KO-IL6-KI (NSGF6) mice were generated using a CRISPR/Cas system. Cas9 mRNA, sgRNAs targeting mouse Il6 and recombinant human IL6 genomic DNA were co-injected into fertilized NOD.Cg-Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl/J (Stock No. 035842) oocytes. Full-length human IL6 sequence replaces exons 1-5 of the mouse Il6 gene. The resulting founders carrying human IL6 were bred to NOD.Cg-Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl/J mice, and were then interbred until all offspring were homozygous for the Il6 targeted mutation. Thereafter, females homozygous for Il6^em1(IL6)Akp, homozygous for Flt3^em1Akp, homozygous for Prkdc^scid, and homozygous for Il2rg^tm1Wjl were bred with males homozygous for Il6^em1(IL6)Akp, homozygous for Flt3^em1Akp, homozygous for Prkdc^scid, and hemizygous for the X-linked Il2rg^tm1Wjl prior to sending female and male mice to The Jackson Laboratory Repository. Upon arrival and recovery, we will maintain our live colony in similar fashion. This strain will be cryopreserved as embryos.

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: Il6^em1(IL6)Akp
Name: endonuclease-mediated mutation 1, Karolina Palucka
MGI accession ID: MGI:7628228
Generation method: Endonuclease-mediated
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: Il6^emX(IL6)Akp
Marker symbol: Il6
Marker name: interleukin 6
Marker synonyms: BSF2; BSF-2; CDF; HGF; HSF; Ifnb2; IFN-beta-2; Il-6; ILg6
Chromosome: 5
Strain of origin: NOD.Cg-Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl/J
Molecular note: Using a CRISPR/Cas9 system, Cas9 mRNA, sgRNAs targeting mouse Il6 and recombinant human IL6 genomic DNA were co-injected into fertilized NOD.Cg-Flt3^em1Akp Prkdc^scid Il2rg^tm1Wjl/J oocyte. Full-length human IL6 sequence replaces exons 1-5 of the mouse Il6 gene.

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.