Triple Knockout mice lack RAG2, CD47, and X-linked IL2RG - making them tolerant of xenotransplantation, resistant to graft versus host disease, and susceptible to HIV.
Of note, a similar strain containing a RAG1 KO, instead of a RAG2 KO, is available as Stock No. 032088.
Kim J. Hasenkrug, National Institutes of Health
Genetic Background | Generation |
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N10+pN3F4
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Null/Knockout) | Il2rg | interleukin 2 receptor, gamma chain |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Null/Knockout) | Cd47 | CD47 antigen (Rh-related antigen, integrin-associated signal transducer) |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Null/Knockout) | Rag2 | recombination activating gene 2 |
Starting at:
$255.00 Domestic price for female 4-week |
510.00 Domestic price for breeder pair |
Triple knockout mice do not express the recombination activating gene 2 (Rag2), the CD47 antigen (CD47) gene, or the X-linked interleukin 2 receptor, gamma chain (Il2rg) gene. These mice are viable and fertile. These mice contain no T cells, B cell, or Natural Killer (NK) cells. The disruption of the Il2rg gene (γc) prevents the development of thymomas and delays the onset of graft versus host disease (GVHD). Recognition of human CD47 by the murine signal-regulatory protein alpha (SIRPα) is required to suppress macrophage-mediated phagocytosis of transplanted human hematopoietic stem cells (HSCs). C57BL/6 mice express a form of the SIRPα receptor that does not recognize human CD47, leading to a lack of tolerization of xenotransplantation. Since phagocytes developed in a CD47-negative environment are tolerized to cells that do not express CD47, disruption of murine CD47 in this C57BL/6 model negates the requirement for CD47-SIRPα signaling and induces tolerance to transplanted human cells.
After transplantation of human bone marrow, liver, and thymus into a similar model, mice develop organized lymphoid tissues and have an intact complement system. Transplantation results in a majority of leukocytes in the peripheral circulation being from human origin. These mice may be good in applications associated with graft versus host disease (GVHD) and virus-specific immune responses.
A targeting vector was designed to replace 0.85kb of the recombination activating gene 2 (Rag2) with a neomycin resistance (neo) cassette and a polyadenylation (polyA) sequence. The construct was electroporated into 129S/SvEv-Gpi1c-derived CCE embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to females. Offspring were subsequently bred to mice bearing the X-linked Il2rgtm1Wjl/J allele (Stock No. 003174). These mice lack exons 4-8 and part of exon 3 of the interleukin 2 receptor, gamma chain (γc). Rag2-/-γc-/- females were bred to B6.129S7-CD47tm1Fpl/J (Stock No. 003173) males. Resulting Rag2+/-γc-/YCD47+/- males were backcrossed to Rag2-/-γc-/-, and mating of F2 Rag2-/-γc-/-CD47+/- females and Rag2-/-γc-/YCD47+/- males produced this Rag2-/-γc-/-CD47-/- (triple knockout; TKO) colony. These mice were bred to C57BL/6J mice (Stock No. 000664) for at least 10 generations.
Allele Name | targeted mutation 1, Warren J Leonard |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | [KO]gammac; CD132-; gammac-; gc-; Il2rgtm1Wjll; IL2Rgammanull |
Gene Symbol and Name | Il2rg, interleukin 2 receptor, gamma chain |
Gene Synonym(s) | |
Site of Expression | Primarily lymphoid cells. |
Strain of Origin | 129S4/SvJae |
Chromosome | X |
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. |
Mutations Made By | Dr. Warren Leonard, NHLBI, NIH |
Allele Name | targeted mutation 1, Frederik P Lindberg |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | CD47-; IAP-; Itgptm1Fpl |
Gene Symbol and Name | Cd47, CD47 antigen (Rh-related antigen, integrin-associated signal transducer) |
Gene Synonym(s) | |
Strain of Origin | 129S7/SvEvBrd-Hprtb-m2 |
Chromosome | 16 |
Molecular Note | A neomycin resistance cassette was inserted into exon 2 of the gene, which encodes the signal peptide cleavage site and the IgV domain. Monoclonal antibodies recognizing the Ig domain failed to detect any CD47 antigen on the surface of erythrocytes, bone marrow cells, or other cells including peripheral blood leukocytes, splenocytes, and fibroblasts from homozygous mutant mice. In addition, no antigen was detected in mutant erythrocyte lysates with polyclonal antisera to the NH2 or COOH-termini. |
Mutations Made By | Dr. Frederik Lindberg, Washington University School of Medicine |
Allele Name | targeted mutation 1, Frederick W Alt |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | RAG-2-; Rag2-; RAG2 KO; Rag2tm1Alt; Rag2tm1Fwe; Rag2M |
Gene Symbol and Name | Rag2, recombination activating gene 2 |
Gene Synonym(s) | |
Strain of Origin | 129S/SvEv-Gpi1c |
Chromosome | 2 |
Molecular Note | A neomycin resistance cassette replaced a 0.85kb segment of the open reading frame, eliminating the encoding of 286 of the 527 amino acids of the protein. |
When maintaining a live colony, these mice are bred as homozygotes for all three alleles. SPF (specific pathogen-free) conditions are recommended.
When using the Triple KO (TKO) mouse strain in a publication, please cite the originating article(s) and include JAX stock #025730 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous for Rag2<tm1Fwa> , Heterozygous for Cd47<tm1Fpl>, Heterozygous females adn Wildtype males for Il2rg<tm1Wjl>/ |
Frozen Mouse Embryo | B6.129S-Rag2<tm1Fwa> Cd47<tm1Fpl> Il2rg<tm1Wjl>/J Frozen Emb | $2595.00 |
Frozen Mouse Embryo | B6.129S-Rag2<tm1Fwa> Cd47<tm1Fpl> Il2rg<tm1Wjl>/J Frozen Emb | $2595.00 |
Frozen Mouse Embryo | B6.129S-Rag2<tm1Fwa> Cd47<tm1Fpl> Il2rg<tm1Wjl>/J Frozen Emb | $3373.50 |
Frozen Mouse Embryo | B6.129S-Rag2<tm1Fwa> Cd47<tm1Fpl> Il2rg<tm1Wjl>/J Frozen Emb | $3373.50 |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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