Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD3EDG/hCD20 mice.
The exons 2-8 of mouse Cd3e gene were replaced by human CD3E exons 2-9 in B-hCD3EDG/hCD20 mice. The exons 1-5 of mouse Cd3d and the exons 1-6 of Cd3g gene were replaced by human CD3D exons 1-5 and CD3G exons 1-7 in B-hCD3EDG/hCD20 mice, respectively. The exons 2-7 of mouse Cd20 gene that encode the full-length protein were replaced by human CD20 exons 2-7 in B-hCD3EDG/hCD20 mice.
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mRNA expression analysis
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Strain specific analysis of CD3DG gene expression in wild type (WT) mice and B-hCD3EDG/hCD20 mice by RT-PCR.
Mouse Cd3d and Cd3g mRNA were detectable only in thymocytes of WT mice (+/+). Human CD3D and CD3G mRNA were detectable only in homozygous B-hCD3EDG/hCD20 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-hCD3EDG/hCD20 mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD3EDG/hCD20 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-hCD3EDG/hCD20 mice (H/H;H/H) but not in WT mice (+/+).
Strain specific CD20 expression analysis in homozygous B-hCD3EDG/hCD20 mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD3EDG/hCD20 mice (H/H;H/H), and analyzed by flow cytometry with species-specific anti-CD20 antibody. Mouse CD20 was detectable in WT mice (+/+). Human CD20 was exclusively detectable in homozygous B-hCD3EDG/hCD20 mice (H/H;H/H) but not in WT mice (+/+).
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Summary
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- mRNA expression analysis:
Mouse Cd3d and Cd3g mRNA were detectable only in thymocytes of WT mice (+/+). Human CD3D and CD3G mRNA were detectable only in homozygous B-hCD3EDG/hCD20 mice (H/H;H/H) but not in WT mice (+/+).
- Protein expression analysis:
Human CD3E was exclusively detectable on T cells of homozygous B-hCD3EDG/hCD20 mice (H/H;H/H) but not in WT mice (+/+), and mouse CD3EDG/hCD20 was detectable only in WT mice (+/+).
Human CD20 was exclusively detectable on B cells of homozygous B-hCD3EDG/hCD20 mice (H/H;H/H) but not in WT mice (+/+), and mouse CD3EDG/hCD20 was detectable only in WT mice (+/+).
