These Cd27 knockout mice have impaired T cell responses to viruses as well as delayed memory cell responses. They may have applications in studies related to immune cell development and peripheral expansion.
Jannie Borst, The Netherlands Cancer Institute
A hygro cassette replaces the entire coding region of the CD27 antigen (Cd27; Tnfrsf7) gene, abolishing gene expression. Homozygotes are viable and fertile. CD27 is a tumor necrosis factor (TNF) receptor family member that in mice is found on hematopoietic stem cells in bone marrow, thymocytes, naive and activated peripheral T cells, subsets of natural killer (NK) cells and activated B cells. CD27 expression can be lost from effector T cells and is absent from IL-17-producing gamma delta T cells. CD27 is not expressed outside of the lymphoid lineage. Upon binding of its ligand, CD70, on activated immune cells, CD27 drives clonal expansion of primed T cells in lymphoid organs and survival of effector T cells in non-lymphoid organs. CD27-deficient mice have a more narrow responder TCR repertoire to antigenic challenge. CD27 also supports effector function of T cells and NK cells and memory formation and function of T cells. CD27-deficient mice have normal conventional T cell and B cell development, but a mildly impaired regulatory T cell development. They have mild to moderate impairments in primary and particularly memory T cell responses to viruses, dependent on the virus type or strain. In the mouse, B cell function is not overtly affected by CD27-deficiency. In the mouse, activated immune cells bearing CD70 can exert feed-back control on hematopoiesis via CD27 on hematopoietic precursors.
A targeting vector was designed to replace the coding region of the CD27 antigen (Cd27) gene with a hygromycin resistance (hygro) cassette. The construct was electroporated into 129P2/OlaHsd-derived E14IB10 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to FVB females to generate a colony of Cd27-/- mice. These mice were backcrossed to C57BL/6 for at least 8 generations (see SNP note below). Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
Please note the Cd27 locus is linked to the Ly49 locus and these mice have retained the 129 Ly49 allotype.
A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. One of the 43 markers throughout the genome were segregating with 129. One marker was segregating with FVB, likely a passenger segment for Cd27 mutation. Also, all 4 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6N genetic background.
|Allele Name||targeted mutation 1, Jannie Borst|
|Allele Type||Targeted (Null/Knockout)|
|Allele Synonym(s)||CD27-; Tnfrsf7-|
|Gene Symbol and Name||Cd27, CD27 antigen|
|Strain of Origin||129P2/OlaHsd|
|Molecular Note||The entire coding region of the gene was replaced with a PGK-hygro cassette via homologous recombination. Immunofluorescence flow cytometry of spleen cells from homozygous mutant animals confirmed the absence of protein expression.|
When maintaining a live colony, homozygous mice may be bred together.
When using the B6.129P2(FVB)-Cd27tm1Jbo/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #019009 in your Materials and Methods section.