Basic Information
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Targeting strategy
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Gene targeting strategy for B-hSIRPA/hCD47/hCD3E mice.
The exons 2-6 of mouse Cd3e gene that encode the extracellular domain were replaced by human CD3E exons 2-7 in B-hSIRPA/hCD47/hCD3E mice. The exon 2 of mouse Cd47 gene that encodes the extracellular domain was replaced by exon2 of human CD47 gene in B-hSIRPA/hCD47/hCD3E mice. The exon 2 of mouse Sirpα gene that encodes the extracellular domain was replaced by exon2 of human SIRPα in B-hSIRPA/hCD47/hCD3E mice.
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Protein expression analysis
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Strain specific CD3E expression analysis in homozygous B-hSIRPA/hCD47/hCD3E mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hSIRPA/hCD47/hCD3E mice (H/H;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-hSIRPA/hCD47/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+).
Strain specific CD47 expression analysis in homozygous B-hSIRPA/hCD47/hCD3E mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hSIRPA/hCD47/hCD3E mice (H/H;H/H;H/H), and analyzed by flow cytometry with species-specific anti-CD47 antibody. Mouse CD47 was detectable in WT mice (+/+). Human CD47 was exclusively detectable in homozygous B-hSIRPA/hCD47/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+).
Strain specific SIRPA expression analysis in homozygous B-hSIRPA/hCD47/hCD3E mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hSIRPA/hCD47/hCD3E mice (H/H;H/H;H/H), and analyzed by flow cytometry with anti-SIRPA antibody. SIRPA was detectable in WT mice (+/+) and homozygous B-hSIRPA/hCD47/hCD3E mice (H/H;H/H;H/H) due to the cross-reactivity of antibodies.
