BAP1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of SK-HEP-1 human liver adenocarcinoma cells featuring targeted disruption of the BAP1 gene. This polyclonal pool provides a heterogeneous loss-of-function model suitable for studying BAP1-dependent processes without clonal isolation. The population-based editing approach enables analysis of gene function across diverse genetic backgrounds, reflecting tumor heterogeneity. These cells allow investigation of BAP1??s roles in DNA repair, chromatin regulation, and cell cycle control within a hepatocellular carcinoma context.
SK-HEP-1 is an adherent epithelial-like cell line derived from ascites of a liver adenocarcinoma patient. Despite its hepatic origin, it exhibits mesenchymal features and high metastatic potential, serving as a well-established in vitro model for aggressive hepatocellular carcinoma. Widely used in cancer biology research, SK-HEP-1 enables studies on invasion, migration, and tumor progression, making it an ideal host for assessing BAP1??s impact on liver cancer cell behavior.
BAP1 encodes a deubiquitinase that specifically removes mono-ubiquitin from histone H2A at lysine 119 (H2AK119ub), thereby regulating transcription of DNA repair and cell cycle genes. It is regulated by ATM/ATR-mediated DNA damage signaling, CDK phosphorylation, and E2F transcription factors. BAP1 forms complexes with ASXL1/2, BRCA1, BARD1, FOXK1/2, HCF-1, and OGT, integrating functions in homologous recombination repair, PRC1 modulation, and chromatin remodeling. Downstream, BAP1 stabilizes HCF-1 and controls expression of targets such as p21/CDKN1A and BAX, promoting cell cycle arrest and apoptosis. Loss of BAP1 impairs homologous recombination, disrupts H2AK119ub dynamics, and fosters genomic instability, contributing to tumorigenesis.
In SK-HEP-1 cells, BAP1 knockout offers a valuable tool to elucidate tumor suppressor mechanisms in hepatocellular carcinoma. Although BAP1 mutations are most frequent in uveal melanoma and mesothelioma, its inactivation may accentuate malignant phenotypes in liver cancer, particularly in mesenchymal contexts. Ablation of BAP1 reveals defects in DNA double-strand break repair, altered histone ubiquitination, and dysregulated apoptosis, all cancer hallmarks. The polyclonal knockout mirrors intratumoral heterogeneity, enabling studies on variable responses to genotoxic stress and epigenetic disruption.
These polyclonal knockout cells are well-suited for functional assays such as Western blotting and RT-qPCR to confirm BAP1 loss, immunofluorescence and flow cytometry for ??H2AX foci and apoptosis detection, and colony formation and homologous recombination reporter assays to assess growth and repair. They can also be employed in xenograft tumor models and drug sensitivity screening to explore BAP1-deficient vulnerabilities. For additional information, please contact Ascent Research.
