Glypican-3 (GPC3) is a heparan sulfate proteoglycan encoded by the GPC3 gene, anchored to the cell membrane via a glycosylphosphatidylinositol (GPI) moiety. It is widely expressed during embryonic development but is expressed at very low or nearly undetectable levels in normal adult liver tissues ^[1-2]. GPC3 is predominantly overexpressed in hepatocellular carcinoma (HCC) and hepatoblastoma, making it an important tumor-associated antigen ^[2-4]. The GPC3-encoded protein participates in the regulation of signaling pathways such as Wnt/β‑catenin, Hedgehog, and YAP, and plays critical roles in tumor cell proliferation, migration, invasion, and modulation of the tumor microenvironment ^[1,3]. Studies have shown that aberrant GPC3 expression is closely associated with poor prognosis, tumor metastasis, and recurrence in HCC ^[4]. Moreover, Gpc3 gene deletion or mutation can lead to neonatal lethality, embryonic overgrowth, and renal cysts ^[5]. Due to its low expression in normal adult tissues and high specificity in HCC, GPC3 has emerged as a key target for targeted therapy and cancer immunotherapy. To date, various GPC3‑directed therapeutic modalities, including CAR‑T cells, CAR‑NK cells, monoclonal antibodies, bispecific antibodies, and antibody‑drug conjugates (ADCs), have been developed for tumor immunotherapy ^[1,3-4,6]. The hGPC3 mouse is a humanized model generated by replacing a partial sequence of exon 1 in the murine Gpc3 gene with the Kozak-Human GPC3 CDS-3'UTR of Mouse Gpc3-WPRE-BGH pA cassette. This model is suitable for evaluating the in vivo efficacy and safety of GPC3-targeted therapeutics, including CAR-T/CAR-NK cells, monoclonal antibodies, bispecific antibodies, and ADCs. Furthermore, it is applicable to research on hepatocellular carcinoma (HCC) pathogenesis, the tumor immune microenvironment, and combination therapy strategies.

Itpr1 is located on chromosome 6 of mice. Nuclease Technology was used to design sgRNA; Itpr1 knockout mice were obtained by applying high-throughput electroporation of fertilized eggs. After sexual maturity, sperm were collected for cryopreservation.

Pigq is located on chromosome 17 of mice. SgRNA and ssDNA will be designed using Nuclease Technology; Pigq conditional knockout mice will be obtained by high-throughput electroporation of fertilized eggs. After sexual maturity, sperm will be collected for cryopreservation.