These mice carry a mutant allele with a phenylalanine substitution for the tyrosine at position 170 in exon 4; homozygotes have IL-2 production and secretion similar to wildtype controls. This mutant mouse strain may be useful in studies of humoral immune response and T cell proliferation.
Jonathan M Green, Washington University School of Medicine
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Not Specified) | Cd28 | CD28 antigen |
Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. These mice carry a mutant allele with a phenylalanine substitution for the tyrosine at position 170 in exon 4 (CD2- Y170F), which encodes proximal tyrosine-based binding motif. The substitution was confirmed by direct sequencing and RT-PCR. Mutant protein expression levels on thymocytes and peripheral T cells are similar to wildtype protein levels in control mice, as assayed by flow cytometry. Homozygotes have IL-2 production and secretion similar to wildtype controls. This mutant mouse strain may be useful in studies of humoral immune response and T cell proliferation.
A targeting vector containing a floxed neo cassette was used to insert a phenylalanine substitution
for the tyrosine at position 170 in exon 4. The construct was electroporated into 129X1/SvJ derived RW-4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to a transgenic EllA-cre strain on the C57BL/6 background to remove the floxed neo selection cassette. The mice were then backcrossed to C57BL/6 for 10 generations. Upon arrival at the MMRRC at The Jackson Laboratory, the mice were crossed to C57BL/6J once to establish the colony.
Allele Name | targeted mutation 2.1, Jonathan M Green |
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Allele Type | Targeted (Not Specified) |
Allele Synonym(s) | CD28-Y170F |
Gene Symbol and Name | Cd28, CD28 antigen |
Gene Synonym(s) | |
Promoter | Cd28, CD28 antigen, mouse, laboratory |
Strain of Origin | 129X1/SvJ |
Chromosome | 1 |
Molecular Note | A TAC to TTC mutation was created in exon 4 to substitute phenylalanine for tyrosine at position 170 (Y170F). A floxed neomycin cassette was inserted upstream of the exon and was subsequently removed by cre-mediated recombination. Expression of the mutant allele in splenocytes and thymocytes was similar to endogenous levels as determined by flow cytometry analysis. This mutation disrupts the motif responsible for interaction with the p85 subunit of PI3-kinase. |
Mutations Made By | Jonathan Green, Washington University School of Medicine |
When maintaining a live colony, these mice can be bred as homozygotes.
When using the CD28-Y170F mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #32040 in your Materials and Methods section.
Facility Barrier Level Descriptions
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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