The GPC3 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population in which the GPC3 gene is disrupted, providing a loss-of-function model for investigating the molecular functions of glypican-3 in human cells. This product is generated by introducing targeted double-strand breaks via CRISPR/Cas9, resulting in a heterogeneous pool of cells with diverse GPC3 mutations.
The host HAP1 cell line is a near-haploid human myeloid leukemia cell line derived from the KBM-7 chronic myeloid leukemia patient. HAP1 cells display an adherent, fibroblast-like morphology and maintain a predominantly haploid karyotype, which is exceptionally advantageous for functional genomics and genetic screens. The haploid genomic context minimizes complexity arising from multiple alleles, allowing straightforward interpretation of knockout phenotypes and making HAP1 an established platform for large-scale CRISPR-based studies.
GPC3 encodes a GPI-anchored heparan sulfate proteoglycan that acts as a co-receptor for Wnt, Hedgehog, and FGF signaling cascades. It directly binds ligands including Wnt3a, Wnt5a, SHH, FGF2, and IGF2, and associates with receptors Frizzled and LRP5/6, thereby modulating pathway activation. GPC3 expression is controlled by transcription factor YY1, DNA methylation, and the aforementioned ligands. Downstream, GPC3 influences ??-catenin/TCF-mediated transcription of proliferative genes such as MYC and cyclin D1, and feedback regulators like AXIN2; it also modulates Hedgehog effectors GLI1 and PTCH1, and YAP/TEAD activity via interaction with CD81. This integrated co-receptor function positions GPC3 centrally in the regulation of cell proliferation and apoptosis, aligning with its roles as an oncofetal protein and tumor suppressor.
Knockout of GPC3 in HAP1 cells disrupts these interconnected signaling networks, enabling precise dissection of pathway crosstalk. The near-haploid genotype eliminates confounding alleles, ensuring that phenotypic changes are directly linked to GPC3 loss. This model is especially pertinent for studying hepatocellular carcinoma, Simpson-Golabi-Behmel syndrome, Wilms tumor, and lung cancer, where GPC3 dysregulation is pathogenic. The polyclonal knockout population provides a heterogeneous allele pool, ideal for functional genomics and drug screening.
This polyclonal knockout cell population supports a range of applications, including functional genomics screens, mechanistic studies of Wnt, Hedgehog, and FGF signaling crosstalk, drug target validation, and developmental biology investigations. Compatible assays include western blotting for ??-catenin and GLI1, RT-qPCR for target genes (e.g., MYC, AXIN2, PTCH1), flow cytometry for apoptosis, proliferation assays, RNA-seq, immunofluorescence, and Wnt reporter assays (TOP/FOP-flash). For additional details, contact Ascent Research.
