Ptprc (CD45) knock-out mice exhibit defective thymocyte maturation, reduced numbers of peripheral T cells, and impaired Igμ-induced B cell proliferation. These mice may be useful for studies of the development and function of T and B lymphocytes.
Dr. Tak Mak, University Health Network/Un of Toronto
The Ptprc (protein tyrosine phosphatase, receptor type, C or CD45) gene encodes a protein tyrosine phosphatase involved in regulating T and B cell antigen receptor signaling. In this CD45 knockout allele, a neomycin cassette was used to disrupt variable exon 6. Immunoflouorescence analysis indicates the CD45 is absent from the surface of all B lymphocytes in bone marrow and spleen. CD45 cell surface expression is absent on CD4- CD8- precursors and immature CD4+ CD8+ thymocytes, however, a fraction of peripheral T cells express CD45 on their cell surface.
Mice homozygous for the allele are viable and fertile, exhibiting no overt phenotype. An impairment in thymocyte maturation results in reduced numbers of peripheral CD8+ T cells, and to a lesser extent CD4+ T cells, despite only a slight reduction in total numbers of thymocytes. Cytoxic T cell response to LCMV infection is absent. B cell development is normal, however, Igμ-induced, but not LPS-induced, B cell proliferation is abolished. These mice may be useful for studies of the development and function of T and B lymphocytes.
A targeting vector containing a neomycin cassette, in reverse orientation, was used to disrupt exon 6. The construct was electroporated into 129/Sv-derived D3NS embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts. The resulting chimeric animals were bred to C57BL/6 to achieve germline transmission. The mice were then backcrossed to C57BL/6 for at least 8 generations. Upon arrival, sperm was cryopreserved. To establish a live colony, an aliquot of frozen sperm is used to fertilize C57BL/6J.
|Allele Name||targeted mutation 1, Tak W Mak|
|Allele Type||Targeted (Null/Knockout)|
|Allele Synonym(s)||cd45-; CD45-exon6-|
|Gene Symbol and Name||Ptprc, protein tyrosine phosphatase, receptor type, C|
|Strain of Origin||129/Sv|
|General Note||ES cell line = D3NS.|
|Molecular Note||Exon 6, which is excised in some isoforms, was deleted and replaced with a neomycin selection cassette. Immunofluorescence analysis, using an antibody raised against all isoforms, of cells isolated from homozygous mutant mice showed an absence of encoded protein in B lymphocytes and low levels of expression in peripheral T cells. Flow cytometric analysis of peripheral T cells confirmed that protein containing the region encoded by exon 6 was absent, while protein containing the region encoded by exon 5 was present at low levels.|
While maintaining a live colony, these mice are bred as homozygotes.
When using the CD45-exon6- mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #42055 in your Materials and Methods section.
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