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Rag2/Il2rg Double Knockout

Constitutive Knockout

Rag2/II2rg Double Constitutive Knockout Mouse Model

Mixed Background

  • Model #
  • Genotype
  • Nomenclature
  • 4111-F
    C57BL/6NTac;B10(Cg)-Rag2tm1Fwa Il2rgtm1Wjl
  • 4111-M
    C57BL/6NTac;B10(Cg)-Rag2tm1Fwa Il2rgtm1Wjl
The common gamma knockout mouse lacks functional receptors for many cytokines including IL-2, IL-4, IL-7, IL-9, and IL-15. As a consequence lymphocyte development is greatly compromised. The mouse lacks natural killer (NK) cells and produces only a small number of T and B cells. To eliminate the residual T and B cells this mouse was crossed to the Rag2 (recombinase activating gene 2) deficient mouse. The double knockout should now lack T cells, B cells, and NK cells. A preliminary analysis of this strain showed no detectable NK1.1+ cells, B220+ B cells or Thy1+ T cells in the spleen. A small number of the 1-3 million cells obtained were CD45 low and probably are cells in the macrophage/monocyte lineage. Functional tests, performed by Drs. Elias Hadaad and Pierre Henkart of NIH, for NK killing activity in the spleen, with or without poly I:C stimulation, were negative. Although these mice still need to be examined for a number of other parameters, they should prove to be EXTREMELY USEFUL for transplanting allogeneic or xenogeneic stem cells which are often rejected by NK cells. The mice will also be of value in combination with the parental Rag2 knockout mice for sorting out the role of NK cells in host resistance to tumors and infectious agents.

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Genetic Background:

C57BL/6J x C57BL/10SgSnAi Background


The common gamma chain (Il2rg) knockout was made by Xiqing Cao and Warren Leonard by deleting exons 4 to 8 and part of exon 3 of the X chromosome linked Il2rg gene in 129S4/SvJae-derived J1 ES cells. The chimeric founders were bred to C57BL/6J mice to N8 and two heterozygous females were sent to NIAID/Taconic on 5/9/97 and bred to a C57BL/10AiTac-Rag2tm1Fwa N13 male. The Rag2 knockout was originally derived from 129 x BALB/c breeding stock created by Drs. Shinkai and Alt. It was crossed 11 times to B10.D2/AiTac and 2 times to C57BL/10AiTac before intracrossing to generate the homozygous Rag2 knockout on a B10 background. F1 Male offspring (from the Il2rg-/y x Rag2+/- intercross) were selected by lack of expression of B220+ cells and bred to C57BL/10AiTac-Rag2tm1Fwa N13 females. The Il2rg+/, Rag2-/- female offspring were identified by PCR genotyping. These females were then mated back to their fathers. Embryos from this mating were embryo transfer-derived into the barrier facility on 12/9/97, and Il2rg+/-, Rag2-/- female and Il2rg-/y, Rag2-/- male mice identified. These were then intracrossed to produce females that were homozygous for both knockout alleles and males that were hemizygous for the Il2rg knockout allele and homozygous for the Rag2 knockout allele. The colony is now maintained by crossing double homozygous female mice with male mice homozygous for the Rag2 knockout allele and hemizygous for the Il2rg knockout allele. These mice have bred true for one generation as Rag2-/-, Il2rg-/-, and H-2Kb+. Taconic has continued backcrossing of the line to C57BL/6NTac. SNP testing shows >99% C57BL/6 background genome.





Initial Publication:

  • Garcia S, DiSanto J, Stockinger B. Following the development of a CD4 T cell response in vivo: from activation to memory formation. Immunity 1999 Aug;11(2):163-71.
  • Greenberg P, Riddell S. Deficient cellular immunity--finding and fixing the defects. Science. 1999 Jul 23;285(5427):546-51. Review.
  • Cao X, Shores EW, Hu-Li J, Anver MR, Kelsall BL, Russell SM, Drago J, Noguchi M, Grinberg A, Bloom ET, et al. Defective lymphoid development in mice lacking expression of the common cytokine receptor gamma chain; Immunity, 1995 Mar;2(3):223-38.
  • Shinkai Y, Rathbun G, Lam KP, Oltz EM, Stewart V, Mendelsohn M, Charron J, Datta M, Young F, Stall AM, and Alt FW. RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangemen; Cell, 1992 Mar 6;68(5):855-67.

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