NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/J

Stock number: 024099
Product name: NRG-SGM3
Strain synonyms: NRGS, NRG-3GS
Research areas: Cancer Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NRGS (NRG-SGM3) mice are NOD.Rag1^-/-;γc^null (NRG) animals expressing human interleukin-3 (IL-3), human granulocyte/macrophage-stimulating factor (GM-CSF) and human Steel factor (SF) from the SGM3 (3GS) triple coinjected transgenes. NRGS mice may be useful for cell or tissue transplantation studies on a background expressing human cytokines, particularly as a model for human lymphohematopoietic cell engraftment studies that require a radioresistant host.

Detailed description

NRGS (NRG-SGM3) mice are NOD.Rag1^-;γc^null (NRG) animals also carrying the SGM3 (3GS) triple coinjected transgenes that express human interleukin-3 (IL-3), human granulocyte/macrophage-stimulating factor (GM-CSF) and human Steel factor (SF).

NRGS females homozygous for both mutations and homozygous for the 3GS triple coinjected transgenes (and NRGS males homozygous for the Rag1^- mutation, hemizygous for the X-linked ;γc^null mutation and homozygous for the 3GS triple coinjected transgenes) are viable and fertile, with no mature T cells, B cells, or natural killer (NK) cells. Like NOD.Rag1^-/-;γc^null mice (NRG; Stock No. 007799), these NRGS mice have the same irradiation/engraftment advantages over NOD.scid ;γc^null (NSG; Stock No. 005557), NOD.Rag1^-/- (Stock No. 003729) and NOD.scid (Stock No. 001303). NRGS differs from NRG only in expression of the 3GS transgenes that, like the 3GS transgenic animals (Stock No. 013062) they are derived from, should constitutively produce 2-4 ng/ml serum levels of the human cytokines IL-3, GM-CSF and SF.

Development

NRGS (NRG-SGM3) mice are Rag1^-;γc^null (NRG) animals also carrying the SGM3 (3GS) triple coinjected transgenes that express human interleukin-3 (IL-3), human granulocyte/macrophage-stimulating factor (GM-CSF) and human Steel factor (SF). The mutations/transgenes are described below.

Creation of the Rag1^- null mutation (Rag1KO or Rag1^tm1Mom) on chromosome 2 and the γc^null mutation (IL2RγcKO or Il2rg^tm1Wjl) on chromosome X is described for Stock No. 007799. NOD-congenic double mutant mice (NOD.Rag1^-/-;γc^null or "NRG" mice) were used for breeding as below.

Dr. C.J. Eaves (British Columbia Cancer Agency) designed three separate transgenes each carrying either the human interleukin-3 gene (IL-3 or IL3), the human granulocyte/macrophage-stimulating factor gene (GM-CSF or CSF2) or the human Steel factor gene (SF, SCF or KITLG). 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 sequence (polyA). Co-injection of the transgenes resulted in mice carrying all three transgenes (collectively called 3/GM/SF, SGM3, 3GS, human SCF/GM-CSF/IL-3 cytokine cassette or Tg(CMV-IL3,CSF2,KITLG)1Eav). Dr. James C. Mulloy (Cincinnati Children's Hospital Medical Center) assembled NOD-congenic 3GS mice also carrying the scid mutation (Prkdc^scid) and γc^null mutation. The resulting NSG-SGM3 (or "NSGS") mice are described as Stock No. 013062. NSGS animals were used for breeding as below.

To generate the NRGS animals (Stock No. 024099), The Jackson Laboratory bred NRG mice (Stock No. 007799) with NSGS mice (Stock No. 013062). Offspring were selectively bred together or to NRG mice to remove the scid mutation. The NRGS colony was thereafter maintained by breeding homozygous females (Rag1^-/-;γc^-/-;3GS^Tg/Tg) with homozygous males (Rag1^-/-;γc^-/Y;3GS^Tg/Tg).

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
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: Rag1^tm1Mom
Name: targeted mutation 1, Peter Mombaerts
MGI accession ID: MGI:1857241
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Rag1
Marker name: recombination activating gene 1
Chromosome: 2
Strain of origin: 129S7/SvEvBrd-Hprt1⁺
Site of expression: expression is seen in bone marrow derived cell lines.
Molecular note: A 1356 bp genomic fragment of the Rag1 gene, encoding the nuclear localization signal and the zinc-finger motif, was replaced by a neomycin cassette. A mutant transcript expressed from this allele was detected by Northern blot in bone marrow derived cell lines from homozygous mice.

Allele

Symbol: Il2rg^tm1Wjl
Name: targeted mutation 1, Warren J Leonard
MGI accession ID: MGI:1857455
Generation method: Targeted
Attributes: Null/Knockout
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.