Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD94/hNKG2A/hNKG2C mice.
The exons 3~6 of mouse Cd94 gene that encode the extracellular domain were replaced by human CD94 exons 3~6 in B-hCD94/hNKG2A/hNKG2C mice;
The exons 2~6 of mouse Nkg2a gene that encode the extracellular domain were replaced by human NKG2A exons 2~6 in B-hCD94/hNKG2A/hNKG2C mice;
The exons 2~5 of mouse Nkg2c gene that encodes the extracelluar domain was replaced by human NKG2C exons 2~6 in B-hCD94/hNKG2A/hNKG2C mice. -
Protein expression analysis
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Strain specific CD94 expression analysis in heterozygous B-hCD94/hNKG2A/hNKG2C mice by flow cytometry. Splenocytes were collected from wild-type mice and heterozygous B-hCD94/hNKG2A/hNKG2C mice and stimulated with anti-CD3ε in vivo (7.5 μg/mice, stimulation for 24 hours, i.p. ), and analyzed by flow cytometry with species-specific anti-CD94 antibody. Human CD94 were exclusively detectable in heterozygous B-hCD94/hNKG2A/hNKG2C but not in wild-type mice.
Strain specific NKG2A expression analysis in heterozygous B-hCD94/hNKG2A/hNKG2C mice by flow cytometry. Splenocytes were collected from wild-type mice and heterozygous B-hCD94/hNKG2A/hNKG2C mice and stimulated with anti-CD3ε in vivo (7.5 μg/mice, stimulation for 24 hours, i.p. ), and analyzed by flow cytometry with species-specific anti-NKG2A antibody. Human NKG2A were exclusively detectable in heterozygous B-hCD94/hNKG2A/hNKG2C but not in wild-type mice.
Strain specific NKG2C expression analysis in heterozygous B-hCD94/hNKG2A/hNKG2C mice by flow cytometry. Splenocytes were collected from wild-type mice and heterozygous B-hCD94/hNKG2A/hNKG2C mice and stimulated with anti-CD3ε in vivo (7.5 μg/mice, stimulation for 24 hours, i.p. ), and analyzed by flow cytometry with species-specific anti-NKG2C antibody. Human NKG2C were exclusively detectable in heterozygous B-hCD94/hNKG2A/hNKG2C mice but not in wild-type mice. -
Summary
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Protein expression analysis:
Human CD94, NKG2A and NKG2C were exclusively detectable on CD8+ T cells and NK cells in splenocytes of heterozygous B-hCD94/hNKG2A/hNKG2C mice but not in wild-type mice, and mouse CD94 and mouse NKG2A were detectable only in wild-type mice. The mouse NKG2C was not tested because there were no useful antibodies available.
