C.B6-Cd3etm2(CD3E)Bcgen Cd3dtm1(CD3D)Bcgen Cd3gtm1(CD3G)Bcgen Tnfrsf9tm1(TNFRSF9)Bcgen/Bcgen • 113254
Product name | B-hCD3EDG/h4-1BB mice(C.B6) |
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Catalog number | 113254 |
Strain name | C.B6-Cd3etm2(CD3E)Bcgen Cd3dtm1(CD3D)Bcgen Cd3gtm1(CD3G)Bcgen Tnfrsf9tm1(TNFRSF9)Bcgen/Bcgen |
Strain background | C57BL/6; BALB/cCrSlcNifdc |
NCBI gene ID | 12501, 12500, 12502, 21942 |
Aliases | CD3epsilon, IMD18, T3E, TCRE;CD3-DELTAELTA, IMD19, T3D, CD3D:CD3-GAMMAAMMA, IMD17, T3G, CD3G;4-1BB, CD137, CDw137, ILA, IMD109 |
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Gene targeting strategy for B-hCD3EDG/h4-1BB mice(C.B6).
Chimeric human CD3EDG were expressed, while mouse Cd3edg were knocked out in B-hCD3EDG/h4-1BB mice(C.B6). The exons 2-7 of the mouse 4-1BB gene, which encode the extracellular domain, were replaced by human counterparts in B-hCD3EDG/h4-1BB mice(C.B6). The genomic region of mouse 4-1BB gene that encodes transmembrane domain and cytoplasmic portion were retained. The promoter, 5’UTR and 3’UTR region of the mouse gene were also retained. The chimeric 4-1BB expression was driven by endogenous mouse 4-1BB promoter, while mouse 4-1BB gene transcription and translation will be disrupted.
Strain specific analysis of CD3E mRNA expression in wild-type BALB/cCrSlcNifdc mice and B-hCD3EDG/h4-1BB mice(C.B6) by RT-PCR. Spleen RNA were isolated from wild-type BALB/cCrSlcNifdc mice (+/+) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CD3E primers. Mouse Cd3e mRNA was only detectable in wild-type mice. Human CD3E mRNA was exclusively detectable in homozygous B-hCD3EDG/h4-1BB mice(C.B6) but not in wild-type mice.
Strain specific analysis of CD3D mRNA expression in wild-type BALB/cCrSlcNifdc mice and B-hCD3EDG/h4-1BB mice(C.B6) by RT-PCR. Spleen RNA were isolated from wild-type BALB/cCrSlcNifdc mice (+/+) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CD3D primers. Mouse Cd3d mRNA was only detectable in wild-type mice. Human CD3D mRNA was exclusively detectable in homozygous B-hCD3EDG/h4-1BB mice(C.B6) but not in wild-type mice.
Strain specific analysis of CD3G mRNA expression in wild-type BALB/cCrSlcNifdc mice and B-hCD3EDG/h4-1BB mice(C.B6) by RT-PCR. Spleen RNA were isolated from wild-type BALB/cCrSlcNifdc mice (+/+) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CD3G primers. Mouse Cd3g mRNA was only detectable in wild-type mice. Human CD3G mRNA was exclusively detectable in homozygous B-hCD3EDG/h4-1BB mice(C.B6) but not in wild-type mice.
Strain specific analysis of 4-1BB mRNA expression in wild-type BALB/cCrSlcNifdc mice and B-hCD3EDG/h4-1BB mice(C.B6) by RT-PCR. Spleen RNA were isolated from wild-type BALB/cCrSlcNifdc mice (+/+) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human 4-1BB primers. Mouse 4-1BB mRNA was only detectable in wild-type mice. Human 4-1BB mRNA was exclusively detectable in homozygous B-hCD3EDG/h4-1BB mice(C.B6) but not in wild-type mice.
Strain specific CD3E expression analysis in wild-type BALB/cCrSlcNifdc mice and homozygous humanized B-hCD3EDG/h4-1BB mice(C.B6) by flow cytometry. Spleen T cells were collected from wild-type BALB/cCrSlcNifdc mice (+/+) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (H/H) (n=3, male, 9-week-old). Protein expression was analyzed with anti-mouse CD3E antibody (Biolegend, 100312) and anti-human CD3E antibody (Biolegend, 562426) by flow cytometry. Mouse CD3E was only detectable in wild-type BALB/cCrSlcNifdc mice. Human CD3E was exclusively detectable in homozygous B-hCD3EDG/h4-1BB mice(C.B6), but not in wild-type BALB/cCrSlcNifdc mice.
Strain specific 4-1BB expression analysis in wild-type BALB/cCrSlcNifdc mice and homozygous humanized B-hCD3EDG/h4-1BB mice(C.B6) by flow cytometry. Spleen T cells were collected from wild-type BALB/cCrSlcNifdc mice (+/+) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (H/H), stimulated with anti-CD3ε in vivo (15 μg/mice for 24 hours, i.p.; BioXcell, BE0001-2, BioXcell, BE0001-1). Protein expression was analyzed with anti-mouse 4-1BB antibody (Biolegend, 106110) and anti-human 4-1BB antibody (Biolegend, 309804) by flow cytometry. Mouse 4-1BB was only detectable in wild-type BALB/cCrSlcNifdc mice. Human 4-1BB was exclusively detectable in homozygous B-hCD3EDG/h4-1BB mice(C.B6), but not in wild-type BALB/cCrSlcNifdc mice.
Strain specific 4-1BB expression analysis in wild-type BALB/cCrSlcNifdc mice and homozygous humanized B-hCD3EDG/h4-1BB mice(C.B6) by flow cytometry. Spleen CD4+ T cells were collected from wild-type BALB/cCrSlcNifdc mice (+/+) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (H/H), stimulated with anti-CD3ε in vivo (15 μg/mice for 24 hours, i.p.; BioXcell, BE0001-2, BioXcell, BE0001-1). Protein expression was analyzed with anti-mouse 4-1BB antibody (Biolegend, 106110) and anti-human 4-1BB antibody (Biolegend, 309804) by flow cytometry. Mouse 4-1BB was only detectable in wild-type BALB/cCrSlcNifdc mice. Human 4-1BB was exclusively detectable in homozygous B-hCD3EDG/h4-1BB mice(C.B6), but not in wild-type BALB/cCrSlcNifdc mice.
Strain specific 4-1BB expression analysis in wild-type BALB/cCrSlcNifdc mice and homozygous humanized B-hCD3EDG/h4-1BB mice(C.B6) by flow cytometry. Spleen CD8+ T cells were collected from wild-type BALB/cCrSlcNifdc mice (+/+) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (H/H), stimulated with anti-CD3ε in vivo (15 μg/mice for 24 hours, i.p.; BioXcell, BE0001-2, BioXcell, BE0001-1). Protein expression was analyzed with anti-mouse 4-1BB antibody (Biolegend, 106110) and anti-human 4-1BB antibody (Biolegend, 309804) by flow cytometry. Mouse 4-1BB was only detectable in wild-type BALB/cCrSlcNifdc mice. Human 4-1BB was exclusively detectable in homozygous B-hCD3EDG/h4-1BB mice(C.B6), but not in wild-type BALB/cCrSlcNifdc mice.
Strain specific 4-1BB expression analysis in wild-type BALB/cCrSlcNifdc mice and homozygous humanized B-hCD3EDG/h4-1BB mice(C.B6) by flow cytometry. Spleen Tregs were collected from wild-type BALB/cCrSlcNifdc mice (+/+) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (H/H), stimulated with anti-CD3ε in vivo (15 μg/mice for 24 hours, i.p.; BioXcell, BE0001-2, BioXcell, BE0001-1). Protein expression was analyzed with anti-mouse 4-1BB antibody (Biolegend, 106110) and anti-human 4-1BB antibody (Biolegend, 309804) by flow cytometry. Mouse 4-1BB was only detectable in wild-type BALB/cCrSlcNifdc mice. Human 4-1BB was exclusively detectable in homozygous B-hCD3EDG/h4-1BB mice(C.B6), but not in wild-type BALB/cCrSlcNifdc mice.
Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type BALB/cCrSlcNifdc mice (male, n=3, 9-week-old) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (male, n=3, 9-week-old). A. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hCD3EDG/h4-1BB mice(C.B6) were similar to those in BALB/cCrSlcNifdc mice. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***p < 0.001.
Frequency of leukocyte subpopulations in blood by flow cytometry. Blood cells were isolated from wild-type BALB/cCrSlcNifdc mice (male, n=3, 9-week-old) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (male, n=3, 9-week-old). A. Flow cytometry analysis of the blood cells was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hCD3EDG/h4-1BB mice(C.B6) were similar to those in BALB/cCrSlcNifdc mice. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***p < 0.001.
Frequency of leukocyte subpopulations in lymph nodes by flow cytometry. Lymph nodes cells were isolated from wild-type BALB/cCrSlcNifdc mice (male, n=3, 9-week-old) and homozygous B-hCD3EDG/h4-1BB mice(C.B6) (male, n=3, 9-week-old). A. Flow cytometry analysis of the lymph nodes cells was performed to assess the frequency of leukocyte subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hCD3EDG/h4-1BB mice(C.B6) were similar to those in BALB/cCrSlcNifdc mice. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***p < 0.001.