Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD3EDG/hBCMA mice. The exons 2-8 of mouse Cd3e gene that encode the full-length protein were replaced by human CD3E exons 2-9 in B-hCD3EDG/hBCMA mice. The exons 1-5 of mouse Cd3d and the exons 1-6 of Cd3g gene that encode the full-length protein were replaced by human CD3D exons 1-5 and CD3G exons 1-7 in B-hCD3EDG/hBCMA mice, respectively. The exon 1 and part of exon 2 encode the extracellular region of mouse Bcma gene were replaced with human BCMA exon 1 and part of exon 2 inB-hCD3EDG/hBCMA mice.
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mRNA expression analysis
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Strain specific analysis of CD3EDG and BCMA gene expression in wild type (WT) mice and B-hCD3EDG/hBCMA mice by RT-PCR. Mouse Cd3edg and Bcma mRNA were detectable only in splenocytes of WT mice (+/+). Human CD3EDG and BCMA mRNA were detectable only in homozygous B-hCD3EDG/hBCMA mice (H/H; H/H) but not in WT mice (+/+). The positive band was confirmed to be correct by sequencing.
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Protein expression analysis
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Strain specific CD3E expression analysis in homozygous B-hCD3EDG/hBCMA mice by flow cytometry. Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD3EDG/hBCMA 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/hBCMA mice (H/H; H/H) but not in WT mice (+/+).
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Summary
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mRNA expression analysis:
- Mouse Cd3edg and Bcma mRNA were detectable only in splenocytes of WT mice (+/+). Human CD3EDG and BCMA mRNA were detectable only in homozygous B-hCD3EDG/hBCMA mice (H/H; H/H) but not in WT mice (+/+). The positive band was confirmed to be correct by sequencing.
Protein expression analysis:
- Human CD3E was exclusively detectable on T cells of homozygous B-hCD3EDG/hBCMA mice (H/H; H/H) but not in WT mice (+/+), and mouse CD3EDG/hBCMA was detectable only in WT mice (+/+).
