C57BL/6-Cd3etm1(CD3E)Bcgen Cd3dtm1(CD3D)Bcgen Cd3gtm1(CD3G)Bcgen Ceacam5tm2(CEACAM5)Bcgen/Bcgen • 114269
| Product name | B-hCD3EDG/hCEACAM5 mice |
|---|---|
| Catalog number | 114269 |
| Strain name | C57BL/6-Cd3etm1(CD3E)Bcgen Cd3dtm1(CD3D)Bcgen Cd3gtm1(CD3G)Bcgen Ceacam5tm2(CEACAM5)Bcgen/Bcgen |
| Strain background | C57BL/6 |
| NCBI gene ID | 916,915,917,1048 (Human) |
| Aliases | T3E; TCRE; IMD18; CD3epsilon; T3D; IMD19; CD3DELTA; CD3-DELTA; T3G; IMD17; CD3GAMMA; CD3-GAMMA; CEA; CD66e |
Gene targeting strategy for B-hCD3EDG/hCEACAM5 mice. The targeting strategy of humanized CD3EDG were undisclosed. The full coding sequences of human CEACAM5 gene, including the promoter, 5’UTR and 3’UTR are inserted into mouse Hipp11 (H11) locus in B-hCD3EDG/hCEACAM5 mice.
CD3E expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCD3EDG/hCEACAM5 mice by flow cytometry. Spleen T cells were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD3EDG/hCEACAM5 mice (H/H; H/H) (Female, 5-week-old, n=1). Protein expression was analyzed with anti-human CD3E antibody (BD Horizon™, 562426) and anti-mouse CD3E antibody (Biolegend, 100312) by flow cytometry. Human CD3E was exclusively detectable in homozygous B-hCD3EDG/hCEACAM5 mice, but not in wild-type C57BL/6JNifdc mice.
Strain specific analysis of CD3D and CD3G mRNA expression in wild-type C57BL/6JNifdc mice and B-hCD3EDG/hCEACAM5 mice by RT-PCR. Spleen RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD3EDG/hCEACAM5 mice (H/H; H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CD3D and CD3G primers. Mouse Cd3d and Cd3g mRNA was only detectable in wild-type mice. Human CD3D and CD3G mRNA was exclusively detectable in homozygous B-hCD3EDG/hCEACAM5 mice but not in wild-type mice.
Strain specific CEACAM5 expression analysis in homozygous B-hCD3EDG/hCEACAM5 mice by flow cytometry. Colon was collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD3EDG/hCEACAM5 mice (H/H, H/H) and analyzed by flow cytometry with species-specific anti-human CD66a/c/e(CEACAM5) Antibody (Biolegend, 342308). Human CEACAM5 was exclusively detectable in homozygous B-hCD3EDG/hCEACAM5 mice but not in wild-type mice.
Western blot analysis of CEACAM5 protein expression in homozygous B-hCD3EDG/hCEACAM5 mice. Various tissue lysates were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD3EDG/hCEACAM5 mice (H/H, H/H), and then analyzed by western blot with human-specific anti-CEACAM5 antibody (Invitrogen, PA5-88533). 40 μg total proteins were loaded for western blotting analysis. Human CEACAM5 were detected in stomach, colon and small intestine tissues in homozygous mice but not wild-type C57BL/6JNifdc mice.
