Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD3E/hHER2 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/hHER2 mice. The exons 2-17 of mouse Erbb2 gene that encode the extracellular domain were replaced by human ERBB2 exons 2-17 in B-hCD3E/hHER2 mice.
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mRNA expression analysis
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Strain specific analysis of CD3E gene expression in wild type (WT) mice and B-hCD3E/hHER2 mice by RT-PCR.
Mouse Cd3e mRNA was detectable only in spleen of WT mice (+/+). Human CD3E mRNA was detectable only in homozygous B-hCD3E/hHER2 mice (H/H;H/H) but not in WT mice (+/+).
Strain specific analysis of HER2 gene expression in wild type (WT) mice and B-hCD3E/hHER2 mice by RT-PCR.
Mouse Erbb2 mRNA was detectable only in liver of WT mice (+/+). Human ERBB2 mRNA was detectable only in homozygous B-hCD3E/hHER2 mice (H/H;H/H) but not in WT mice (+/+).
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Protein expression analysis
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Strain specific CD3E expression analysis in homozygous B-hCD3E/hHER2 mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD3E/hHER2 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/hHER2 mice (H/H;H/H) but not in WT mice (+/+).
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IHC analysis of HER2 expression
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Immunohistochemical (IHC) analysis of HER2 expression in homozygous B-hCD3E/hHER2 mice.
The mammary gland tissues were collected from WT and homozygous B-hHER2 mice (H/H; H/H) and analyzed by IHC with anti-Her2 antibody. HER2 was detectable in WT mice and homozygous B-hHER2 mice due to the cross-reactivity of the antibody. The arrow indicates tissue cells with positive HER2 staining (brown).
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Summary
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mRNA expression analysis:
Mouse Cd3e mRNA was detectable only in spleen of WT mice (+/+). Human CD3E mRNA was detectable only in homozygous B-hCD3E/hHER2 mice (H/H;H/H) but not in WT mice (+/+).
Mouse Erbb2 mRNA was detectable only in liver of WT mice (+/+). Human ERBB2 mRNA was detectable only in homozygous B-hCD3E/hHER2 mice (H/H;H/H) but not in WT mice (+/+).
Protein expression analysis:
Human CD3E was exclusively detectable on T cells of homozygous B-hCD3E/hHER2 mice (H/H;H/H) but not in WT mice (+/+), and mouse CD3E was detectable only in WT mice (+/+).
HER2 was detectable in mammary gland tissues of WT mice and homozygous B-hHER2 mice due to the cross-reactivity of the antibody.
