C57BL/6-Tslptm1(TSLP)Bcgen Crlf2tm2(CRLF2)Bcgen Il7rtm1(IL7R)Bcgen Tnfrsf4tm1(TNFRSF4)Bcgen Tnfsf4tm1(TNFSF4)Bcgen/Bcgen • 113916
| Product name | B-hTSLP/hTSLPR plus/hOX40/hOX40L mice |
|---|---|
| Catalog number | 113916 |
| Strain name | C57BL/6-Tslptm1(TSLP)Bcgen Crlf2tm2(CRLF2)Bcgen Il7rtm1(IL7R)Bcgen Tnfrsf4tm1(TNFRSF4)Bcgen Tnfsf4tm1(TNFSF4)Bcgen/Bcgen |
| Strain background | C57BL/6 |
| NCBI gene ID | 85480,64109,3575,7293,7292 (Human) |
| Aliases | CRL2; TSLPR; CRLF2Y; ILRA; CD127; IL7RA; CDW127; IMD104; sIL-7R; lnc-IL7R; IL7Ralpha; IL-7Ralpha; IL-7R-alpha; OX40; ACT35; CD134; IMD16; TXGP1L; GP34; CD252; OX4OL; TXGP1; CD134L; OX-40L; TNLG2B |
Gene targeting strategy for B-hTSLP/hTSLPR plus/hOX40/hOX40L mice.
The exons 1-5 of mouse Tslp gene that encode the whole molecule (ATG to STOP codon) were replaced by human counterparts in B-hTSLP/hTSLPR plus/hOX40/hOX40L mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human TSLP expression is driven by endogenous mouse Tslp promoter, while mouse Tslp gene transcription and translation will be disrupted.
A chimeric CDS that encodes human TSLPR extracellular domain, transmembrane and mouse TSLPR cytoplasmic domain, followed by mouse 3’UTR-STOP is inserted right after mouse Tslpr signal peptide sequence to replace part of the exon 2 of mouse Tslpr gene. The chimeric TSLPR protein expression will be driven by endogenous mouse Tslpr promoter, while mouse Tslpr gene transcription and translation will be disrupted.
The exons 1~6 of mouse Il7r gene that encode the extracellular region were replaced by human IL7R exons 1~6 in B-hTSLP/hTSLPR plus/hOX40/hOX40L mice.
The exons 1-5 of mouse Ox40 gene that encode the extracellular domain were replaced by human OX40 exons 1-5 in B-hTSLP/hTSLPR plus/hOX40/hOX40L mice.
The exons 2-3 of mouse Ox40l gene that encode the extracellular region were replaced by human OX40L exons 2-3 in B-hTSLP/hTSLPR plus/hOX40/hOX40L mice.
B-hTSLP/hTSLPR plus/hOX40/hOX40L mice were derived from mating B-hTSLP/hTSLPR mice plus (112744) and B-hOX40/hOX40L mice (120543).
Strain specific TSLP expression analysis in homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice by ELISA. Calcipotriol (MC903) was dissolved in ethanol and applied on ears of wild-type C57BL/6JNifdc mice and homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice for 7 days. The ear grinding supernatant collected from the two strains of mouse were analyzed by ELISA (anti-mouse TSLP antibody: Biolegend, 434107; anti-human TSLP antibody: Biolegend, 434207). Mouse TSLP was only detectable in wild-type C57BL/6JNifdc mice. Human TSLP was only detectable in homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice but not in wild-type mice. ND: not detectable.
Strain specific TSLPR expression analysis in homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice by flow cytometry. Bone marrow derived dendritic cells (BMDCs) were produced by culturing the bone marrow from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice (H/H;H/H;H/H;H/H;H/H), which were induced with FLT3L (acrobiosystems, FLL-H5218) for 6 days and respectively stimulated with 1 μg/mL LPS in vitro for 24 h. Protein expression was analyzed with anti-mouse TSLPR antibody (Biolegend, 151805) and anti-human TSLPR antibody (Biolegend, 322906) by flow cytometry. Mouse TSLPR was detectable in wild-type C57BL/6JNifdc mice. Human TSLPR was detectable in homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice. WT：C57BL/6JNifdc, HO: homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice.
Strain specific IL7R expression analysis in homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice(+/+) and homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice (H/H;H/H;H/H;H/H;H/H), protein expression was analyzed by flow cytometry with species-specific anti-mouse IL7R antibody (Biolegend, 135011) and anti-human IL7R antibody (Biolegend, 351303). Mouse IL7R was detectable on T cells, DCs and NK cells of wild-type C57BL/6JNifdc mice. Human IL7R was detectable on T cells, DCs and NK cells of homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice. WT：C57BL/6JNifdc, HO: homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice.
Strain specific IL7R expression analysis in homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice(+/+) and homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice (H/H;H/H;H/H;H/H;H/H) stimulated with PBS or anti-mouse CD3ε antibody in vivo for 24 h, protein expression was analyzed by flow cytometry with species-specific anti-mouse OX40 antibody (Biolegend, 119414) and anti-human OX40 antibody (Biolegend, 350004). Mouse OX40 was detectable on T cells of wild-type C57BL/6JNifdc mice. Human OX40 was detectable on T cells of homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice. WT：C57BL/6JNifdc, HO: homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice.
Strain specific OX40L expression analysis in homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice by flow cytometry. Bone marrow derived dendritic cells (BMDCs) were produced by culturing the bone marrow from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice (H/H;H/H;H/H;H/H;H/H), which were stimulated with LPS in vitro for 24 h. Protein expression was analyzed with anti-mouse OX40L antibody (Biolegend, 108814) and anti-human OX40L antibody (BD, 563766) by flow cytometry. Mouse OX40L was detectable in wild-type C57BL/6JNifdc mice. Human OX40L was detectable in homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice. WT：C57BL/6JNifdc, HO: homozygous B-hTSLP/hTSLPR plus/hOX40/hOX40L mice.
