Basic Information
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Targeting strategy
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Gene targeting strategy for B-hB2M/HLA-E mice.
The exons 1-5 of mouse H2-T23 gene that encode extracellular domain and transmembrane domain is replaced by human HLA-E gene counterparts fused to human B2M through a flexibe (G4S)3 linker in B-hB2M/HLA-E mice. The genomic region of mouse H2-T23 gene that encodes cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric HLA-E expression is driven by endogenous mouse H2-T23 promoter, while mouse H2-T23 gene transcription and translation will be disrupted.
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Protein expression analysis in Blood
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Strain specific HLA-E and Qa-1(b) expression analysis in homozygous B-hB2M/HLA-E mice by flow cytometry. Blood cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hB2M/HLA-E mice (H/H) stimulated with mouse IFN gamma (50 μg, qdx3, i.p.) (Novoprotein, C746) in vivo. Protein expression was analyzed with anti-Qa-1(b) antibody (BD, 566640) and anti-HLA-E antibody (Biolegend, 342606) by flow cytometry. HLA-E was detectable in homozygous mice only. Qa-1(b) was detectable in C57BL/6 only.
Strain specific HLA-E and Qa-1(b) expression analysis in homozygous B-hB2M/HLA-E mice by flow cytometry. Blood cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hB2M/HLA-E mice (H/H) stimulated with mouse IFN gamma (50 μg, qdx3, i.p.) (Novoprotein, C746) in vivo. Protein expression was analyzed with anti-Qa-1(b) antibody (BD, 566640) and anti-HLA-E antibody (Biolegend, 342606) by flow cytometry. HLA-E was detectable in homozygous mice only. Qa-1(b) was detectable in C57BL/6 only.
Strain specific B2M expression analysis in homozygous B-hB2M/HLA-E mice by flow cytometry. Blood cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hB2M/HLA-E mice (H/H) stimulated with mouse IFN gamma (50 μg, qdx3, i.p.) (Novoprotein, C746) in vivo. Protein expression was analyzed with anti-mouse B2m antibody (Biolegend, 154504) and anti-human B2M antibody (Biolegend, 316311) by flow cytometry. Human B2M was detectable in homozygous mice only. Mouse B2m was detectable in homozygous mice and C57BL/6.
Strain specific B2M expression analysis in homozygous B-hB2M/HLA-E mice by flow cytometry. Blood cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hB2M/HLA-E mice (H/H) stimulated with mouse IFN gamma (50 μg, qdx3, i.p.) (Novoprotein, C746) in vivo. Protein expression was analyzed with anti-mouse B2m antibody (Biolegend, 154504) and anti-human B2M antibody (Biolegend, 316311) by flow cytometry. Human B2M was detectable in homozygous mice only. Mouse B2m was detectable in homozygous mice and C57BL/6.
