C57BL/6-Tnfsf8tm2(TNFSF8)Bcgen/Bcgen • 113224
Gene targeting strategy for B-hCD30L mice. The exons 1-4 of mouse Cd30l gene that encode extracellular domain were replaced by human counterparts in B-hCD30L mice. The genomic region of mouse Cd30l gene that encodes transmembrane domain and cytoplasmic portion was retained. The promoter and 5’UTR region of the mouse gene were also retained. The 3’UTR region of the mouse gene are replaced by human counterparts. The CD30L expression was driven by endogenous mouse Cd30l promoter, while mouse Cd30l gene transcription and translation will be disrupted.
Species specific analysis of CD30L gene expression in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCD30L mice by RT-PCR. Spleen was collected from wild-type C57BL/6JNifdc mice(+/+) and homozygous B-hCD30L mice(H/H). Mouse Cd30l mRNA was only detectable in wild-type C57BL/6JNifdc mice. Human CD30L mRNA was only detectable only in homozygous B-hCD30L mice, but not in wild-type C57BL/6JNidfc mice.
Strain specific CD30L expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCD30L mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice(+/+) and homozygous B-hCD30L mice(H/H). Splenocytes are incubated in a medium containing Cell Activation Cocktail (with Brefeldin A) (biolegend, 423303) before analysis of CD30L surface expression on gated T cells. Protein expression was analyzed with anti-mouse CD30L antibody(Biolegend, 106405) and anti-human CD30L antibody(RD, FAB1028A) by flow cytometry. Mouse CD30L was exclusively detectable in wild-type C57BL/6JNifdc mice. Human CD30L was exclusively detectable in homozygous B-hCD30L mice, but not in wild-type C57BL/6JNifdc mice.