• GPC3 is a member of the heparan sulfate proteoglycan family anchored to the cell membrane. GPC3 activates the classical Wnt/β-catenin pathway in hepatocellular carcinoma to manifest an oncogene role.
• A chimeric CDS that encodes human GPC3 extracellular domain, followed by mouse 3’UTR-STOP is inserted right after mouse Gpc3 signal peptide to replace partial exon 3 of mouse Gpc3 gene. The chimeric GPC3 protein expression will be driven by endogenous mouse Gpc3 promoter, while mouse Gpc3 gene transcription and translation will be disrupted.
• Mouse Gpc3 mRNA was only detectable in wild-type B-NDG mice. Human GPC3 mRNA was only detectable in homozygous B-NDG hGPC3 mice.
• Application: This product is used for validation of the efficacy of different antibody drugs and preclinical drug evaluation.

Gene targeting strategy for B-NDG hGPC3 mice. A chimeric CDS that encodes human GPC3 extracellular domain, followed by mouse 3’UTR-STOP is inserted right after mouse Gpc3 signal peptide to replace partial exon 3 of mouse Gpc3 gene. The chimeric GPC3 protein expression will be driven by endogenous mouse Gpc3 promoter, while mouse Gpc3 gene transcription and translation will be disrupted.

Species specific analysis of GPC3 gene expression in wild-type B-NDG mice and homozygous humanized B-NDG hGPC3 mice by RT-PCR. Kidney was collected from wild-type B-NDG mice (+/+) and homozygous B-NDG hGPC3 mice (H/H). Mouse Gpc3 mRNA was detectable in wild-type B-NDG mice but not in homozygous B-NDG hGPC3 mice. Human GPC3 mRNA was detectable in homozygous B-NDG hGPC3 mice but not in wild-type B-NDG mice.

Immunohistochemical (IHC) analysis of GPC3 protein expression in wild-type B-NDG mice and B-NDG hGPC3 mice. The liver and fetal liver were collected from B-NDG mice (+/+) and homozygous B-NDG hGPC3 mice (H/H) and analyzed by IHC with anti-GPC3 antibody (GC33-hIgG1 analog (in-house). Human GPC3 was detectable in fetal liver of homozygous B-NDG hGPC3 mice. The arrow indicates tissue cells with positive GPC3 staining (brown).