B-hCD3E/hGPRC5D mice
| Strain Name | C57BL/6-Cd3etm2(CD3E)Bcgen Gprc5dtm1(GPRC5D)Bcgen/Bcgen | Common Name | B-hCD3E/hGPRC5D mice |
| Background | C57BL/6 | Catalog number | 112601 |
| Aliases |
CD3E, CD3epsilon, IMD18, T3E, TCRE, CD3 epsilon subunit of T-cell receptor complex; GPRC5D, G protein-coupled receptor class C group 5 member D |
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模型验证
Targeting strategy
Gene targeting strategy for B-hCD3E/hGPRC5D mice. The exons 2-6 of mouse Cd3e gene that encode the extracellular domain were replaced by human CD3E exons 2-7 in B-hCD3E/hGPRC5D mice. The exons 2-4 of mouse Gprc5d gene that encode the full-length protein were replaced by human GPRC5D exons 1-3 in B-hCD3E/hGPRC5D mice.
Strain specific CD3E expression analysis in homozygous B-hCD3E/hGPRC5D mice by flow cytometry. Splenocytes were collected from 4 week old wild type mice (+/+) and homozygous B-hCD3E/hGPRC5D mice (H/H; H/H). Protein expression was analyzed with anti-mouse CD3e antibody (Biolegend, 100312) and anti-human CD3e antibody (BD Horizon™, 562426) by flow cytometry. Mouse CD3E was detectable in wild type mice. Human CD3E was exclusively detectable in homozygous B-hCD3E/hGPRC5D mice but not in wild type mice.
GPRC5D expression analysis in homozygous B-hCD3E/hGPRC5D mice by western blot. Spleen, liver and testis were collected from 4 week old wild type mice (+/+) and homozygous B-hCD3E/hGPRC5D mice (H/H; H/H) analyzed by western blot with anti-GPRC5D antibody (abcam, ab55044). GPRC5D was detectable in wild type mice and homozygous B-hCD3E/hGPRC5D mice due to the cross-reactivity of antibody.
Strain specific analysis of GPRC5D mRNA expression in wild-type C57BL/6 mice and B-hCD3E/hGPRC5D mice by RT-PCR. Spleen RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hCD3E/hGPRC5D mice (H/H; H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human GPRC5D primers. Mouse Gprc5d mRNA was detectable in wild-type C57BL/6 mice. Human GPRC5D mRNA was detectable only in homozygous B-hCD3E/hGPRC5D mice but not in wild-type mice.
Complete blood count (CBC). Blood from C57BL/6 and B-hCD3E/hGPRC5D mice (n=8, 8 week old) was collected and analyzed for CBC. There were no differences among any measurement between C57BL/6 and B-hCD3E/hGPRC5D mice, indicating that CD3E and GPRC5D humanized do not change blood cell composition and morphology. Values are expressed as mean ± SEM.
Blood biochemistry tests of B-hCD3E/hGPRC5D mice. Serum from the C57BL/6 and B-hCD3E/hGPRC5D mice (n=8, 8 week old) was collected and analyzed for levels of ALT and AST. There were no differences on either measurement between C57BL/6 and B-hCD3E/hGPRC5D mice, indicating that CD3E and GPRC5D humanized do not change ALT and AST levels or health of liver. Values are expressed as mean ± SEM.
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