Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD3E/hB7-H3 mice.
The exons 2-6 of mouse Cd3e gene that encode the extracellular domain were replaced by human CD3E exons 2-7 in B-hCD3E/hB7-H3 mice.
The exon 3-4 of mouse Cd276 gene that encode the extracellular domain were replaced by human CD276 exon 3-6 in B-hCD3E/hB7-H3 mice.
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Protein expression analysis
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Strain specific CD3E expression analysis in homozygous B-hCD3E/hB7-H3 mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD3E/hB7-H3 mice (H/H; H/H), and analyzed by flow cytometry with species-specific anti-CD3E antibody. Mouse CD3E was detectable in WT mice (+/+). Human CD3E was exclusively detectable in homozygous B-hCD3E/hB7-H3 mice (H/H; H/H) but not in WT mice (+/+).
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mRNA and protein expression analysis
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Strain specific analysis of B7-H3 expression in wild type (WT) mice and B-hCD3E/hB7-H3 mice by RT-PCR and WB.
Lung were collected from wild type (WT) mice (+/+) and homozygous B-hCD3E/hB7-H3 mice (H/H; H/H), and analyzed by RT-PCR and western blot with anti-human B7-H3 antibody. Mouse B7-h3 mRNA was detectable only in lung of WT mice (+/+). Human B7-H3 mRNA was detectable only in homozygous B-hCD3E/hB7-H3 mice (H/H; H/H) but not in WT mice (+/+). The positive band was confirmed to be correct by sequencing. Human B7-H3 was exclusively detectable in homozygous B-hCD3E/hB7-H3 mice (H/H; H/H).
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Summary
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- mRNA expression analysis:
Mouse B7-h3 mRNA was detectable only in lung of WT mice (+/+). Human B7-H3 mRNA was detectable only in homozygous B-hCD3E/hB7-H3 mice (H/H; H/H) but not in WT mice (+/+). The positive band was confirmed to be correct by sequencing.
- Protein expression analysis:
Mouse CD3E was detectable on T cells of WT mice (+/+). Human CD3E was exclusively detectable in homozygous B-hCD3E/hB7-H3 mice (H/H; H/H) but not in WT mice (+/+).
Human B7-H3 was exclusively detectable in homozygous B-hCD3E/hB7-H3 mice (H/H; H/H).
