RAG2 knock-out mice have impaired T and B-cell development and may be useful in the study of hematopoietic and immune system defects, cancer, toxicology, and xenograft/transplant studies.
IMR Colony, The Jackson Laboratory
These RAG-2del mutant mice harbor a pan deletion of exon 3 of the targeted locus. Homozygotes (RAG-2del/del) are viable and fertile, with pan deletion of the entire RAG-2 protein coding region. Homozygous mice may be expected to have the same knockout phenotype as other RAG-2 null mutants or similarly created RAG-2 exon 3 pan-deleted mutants; with hematopoietic and immune system defects including arrested B cell and T cell development at the pro-B and the pro-T cell stages, respectively. These RAG-2del mice may be useful in studying the role of RAG-2 in B cell and T cell development (including cancer and toxicology research as a xenograft/transplant host), T and B cell receptor (V(D)J) recombination, hematopoiesis, hematology, immunology, and inflammation research.
In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. It should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.
A targeting vector was designed to insert a loxP-flanked neomycin resistance gene on one side, and a third loxP site on the other side of exon 3 of the recombination activating gene 2 (Rag2) locus. This construct was electroporated into C57BL/6-derived Bruce-4 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a cre-expressing vector to remove the selection cassette. ES cells in which the neomycin resistance cassette was deleted (leaving a single loxP site upstream, and a single loxP downstream of exon 3) were injected into recipient blastocysts. Chimeric males were bred with C57BL/6 females to generate RAG-2fl mice. The RAG-2fl mice were bred with transgenic mice on mixed genetic background, and then bred with mutant mice on a C57BL/6 genetic background for 8 generations (see SNP note below). The donating investigator reports that both the transgene and other mutant gene were bred out of the RAG-2fl mice prior to arrival at The Jackson Laboratory (as Stock No. 008309). Upon arrival, some RAG-2fl males were bred to cre-deleter females (Zp3-Cre transgenic mice on a C57BL/6 genetic background; Stock No. 003651), and then F1 males were bred back to B6.Zp3-Cre females. The resulting RAG-2del (exon 3 deleted) offspring were bred to C57BL/6J inbred mice (selecting for the RAG-2del mutation and against the cre transgene) for at least 1 generation to generate this RAG-2 "knockout" strain (Stock No. 008449).
|Allele Name||targeted mutation 1.1, University of Cologne|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Rag2, recombination activating gene 2|
|Strain of Origin||B6.Cg-Thy1a|
|Molecular Note||This allele is a derivative of Rag2tm1Cgn in which the sequences flanked by loxP sites were deleted in vivo by crossing mice carrying this allele to a deleter strain.|
|Mutations Made By|| |
Klaus Rajewsky, Max Delbruck Centre for Molecular Medicine
When maintaining a live colony, homozygous RAG2 KO mice may be bred together.
When using the RAG2 KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #008449 in your Materials and Methods section.
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