Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD3EDG/hTROP2 mice.
The exons 2-8 of mouse Cd3e gene were replaced by human CD3E exons 2-9 in B-hCD3EDG/hTROP2 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/hTROP2 mice, respectively. The exon 1 of mouse Trop2 gene that encodes the full-length protein was replaced by human TROP2 exon 1 in B-hCD3EDG/hTROP2 mice.
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mRNA expression analysis
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Strain specific analysis of CD3D and CD3G gene expression in wild type (WT) mice and B-hCD3EDG/hTROP2 mice by RT-PCR.
Mouse Cd3d and Cd3g mRNA were detectable only in splenocytes of WT mice (+/+). Chimeric CD3D and CD3G mRNA were detectable only in homozygous B-hCD3EDG/hTROP2 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/hTROP2 mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD3EDG/hTROP2 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/hTROP2 mice (H/H;H/H) but not in WT mice (+/+).
Strain specific TROP2 expression analysis in homozygous B-hCD3EDG/hTROP2 mice by western blot.
Skin tissue was collected from wild type (WT) mice (+/+) and homozygous B-hCD3EDG/hTROP2 mice (H/H;H/H), and analyzed by western blot with anti-TROP2 antibody. Mouse TROP2 was detectable in WT mice (+/+). Human TROP2 was detectable in homozygous B-hCD3EDG/hTROP2 mice (H/H;H/H) and B-hTROP2 MC38 cell line.
