C57BL/6-Pdcd1tm3(PDCD1)Bcgen Cd274tm1(CD274)Bcgen Vegfatm1(VEGFA)Bcgen/Bcgen • 114019
| Product name | B-hPD-1 plus/hPD-L1/hVEGFA mice |
|---|---|
| Catalog number | 114019 |
| Strain name | C57BL/6-Pdcd1tm3(PDCD1)Bcgen Cd274tm1(CD274)Bcgen Vegfatm1(VEGFA)Bcgen/Bcgen |
| Strain background | C57BL/6 |
| NCBI gene ID | 5133,29126,7422 (Human) |
| Aliases | PD1; PD-1; CD279; SLEB2; hPD-1; hPD-l; hSLE1; ADMIO4; AIMTBS; B7-H; B7H1; PDL1; PD-L1; ADMIO5; hPD-L1; PDCD1L1; PDCD1LG1; VPF; VEGF; MVCD1; L-VEGF |
Gene targeting strategy for B-hPD-1 plus/hPD-L1/hVEGFA mice.
A chimeric CDS that encodes human PDCD1 extracellular domain, mouse Pdcd1 transmembrane and cytoplasmic domain, followed by WPRE-pA is inserted right after mouse Pdcd1 ATG to replace the exon 1 of mouse Pdcd1 gene. The chimeric PDCD1 protein expression will be driven by endogenous mouse Pdcd1 promoter, while mouse Pdcd1 gene transcription and translation will be disrupted.
The exon 3 of mouse Cd274 gene that encodes the IgV domain was replaced by human CD274 exon 3 in B-hPD-1 plus/hPD-L1/hVEGFA mice.
The exons 1-8 of mouse Vegfa gene that encode the full-length protein were replaced by human VEGFA exons 1-8 in B-hPD-1 plus/hPD-L1/hVEGFA mice.
Note: The B-hPD-1 plus/hPD-L1/hVEGFA three knock-in model, was developed by breeding the B-hPD-1 plus mice, the B-hPD-L1 mice and the B-hVEGFA mice.
Strain specific PD-1 expression analysis in wild-type C57BL/6 and homozygous B-hPD-1 plus/hPD-L1/hVEGFA mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1 plus/hPD-L1/hVEGFA mice (H/H) after stimulated with anti-mouse CD3ε antibody (7.5 μg, i.p.) in vivo for 24 hrs (female, 6-week-old, n=1) or not. Protein expression was analyzed with anti-mouse PD-1 antibody (Biolegend, 109104) and anti-human PD-1 antibody (Biolegend, 329908) by flow cytometry. Mouse PD-1 were detectable in wild-type C57BL/6 mice. Human PD-1 were exclusively detectable in homozygous B-hPD-1 plus/hPD-L1/hVEGFA mice but not in wild-type mice.
Strain specific PD-L1 expression analysis in wild-type C57BL/6 and homozygous B-hPD-1 plus/hPD-L1/hVEGFA mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1 plus/hPD-L1/hVEGFA mice (H/H) after stimulated with anti-mouse CD3ε antibody (7.5 μg, i.p.) in vivo for 24 hrs (female, 6-week-old, n=1) or not. Protein expression was analyzed with anti-mouse PD-L1 antibody (Biolegend, 124312) and anti-human PD-L1 antibody (Biolegend, 329706) by flow cytometry. Mouse PD-L1 were detectable in wild-type C57BL/6 mice. Human PD-L1 were exclusively detectable in homozygous B-hPD-1 plus/hPD-L1/hVEGFA mice but not in wild-type mice.
Strain specific VEGFA expression analysis in wild-type C57BL/6 mice and homozygous B-hPD-1 plus/hPD-L1/hVEGFA mice by ELISA. Lung homogenates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1/hPD-L1 plus/hVEGFA mice (H/H; H/H; H/H). Expression level of mouse and human VEGFA were analyzed by ELISA (anti-mouse VEGFA antibody: R&D, MMV00; anti-human VEGFA antibody: R&D, DVE00). Mouse VEGFA was detectable in wild-type mice. Human VEGFA was exclusively detectable in homozygous B-hPD-1 plus/hPD-L1/hVEGFA mice but not in wild-type mice.
