Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCRTAM mice.
The exons 1~8 of mouse Crtam gene that encode the extracellular domain was replaced by human CRTAM exons 1~8 in B-hCRTAM mice.
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Protein expression analysis in CD8+ T cells of spleen
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Strain specific CRTAM expression analysis in homozygous B-hCRTAM mice by flow cytometry. CD8+ T cells were sorted from the splenocytes in wild-type C57BL/6 mice (+/+) and homozygous B-hCRTAM mice (H/H) and stimulated with anti-mCD3e in vitro (2 μg/mL, stimulation for 36 hours) and anti-mCD28 in vitro (5 μg/mL, stimulation for 36 hours), and analyzed by flow cytometry with species-specific anti-CRTAM antibody. Mouse CRTAM was exclusively detectable in wild-type mice. Human CRTAM was exclusively detectable in homozygous B-hCRTAM mice but not in wild-type mice.
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mRNA expression analysis
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Strain specific analysis of CRTAM mRNA expression in wild-type C57BL/6 mice and B-hCRTAM mice by RT-PCR. Spleen RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hCRTAM mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CRTAM primers. Mouse Crtam mRNA were detectable only in wild-type C57BL/6 mice. Human CRTAM mRNA was detectable only in homozygous B-hCRTAM mice but not in wild-type mice.
