The BAP1 Knockout Ca Ski Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population of human Ca Ski cells, in which the gene encoding the BRCA1-associated protein-1 (BAP1) deubiquitinase has been disrupted. This bulk knockout population provides a genetically heterogeneous, loss-of-function model of the tumor suppressor BAP1 in a cervical cancer background, avoiding clonal artifacts and enabling robust assessment of BAP1-dependent phenotypes. The genome editing strategy introduces targeted gene disruption without transgene integration, making this product suitable for studies requiring stable BAP1 deficiency.
The Ca Ski host cell line is an adherent epithelial cell line derived from a metastasis of a cervical epidermoid carcinoma and is widely used as an HPV16-positive cervical cancer model. These cells retain characteristics of squamous cell carcinoma and are commonly employed in investigations of human papillomavirus (HPV)-driven oncogenesis, DNA damage response, and tumor suppressor pathways. Their endogenous expression of HPV E6 and E7 oncoproteins offers a relevant context for examining the interplay between viral transformation and host tumor suppressors such as BAP1.
BAP1 functions as a nuclear deubiquitinase that removes monoubiquitin from histone H2A at lysine 119, counteracting Polycomb-mediated gene repression. It forms the catalytic core of the PR-DUB complex with ASXL1/2 and interacts with BRCA1, BARD1, HCF-1, and OGT to coordinate DNA repair, chromatin remodeling, and ferroptosis. Upon DNA double-strand breaks, ATM/ATR kinases activate BAP1, promoting deubiquitination of H2AK119ub and enabling expression of the INK4A/ARF locus while modulating YAP/TAZ through Hippo signaling. Loss of BAP1 results in sustained H2AK119ub, aberrant silencing of Polycomb targets, and oncogenic YAP/TAZ activation.
In the Ca Ski background, BAP1 knockout is particularly relevant for dissecting the tumor suppressive mechanisms counteracting HPV-driven cervical carcinogenesis. HPV E6 and E7 oncoproteins disrupt p53 and retinoblastoma (Rb) pathways, while BAP1 further safeguards genome stability and epigenetic fidelity. The polyclonal knockout population enables investigation of BAP1??s role in modulating HPV oncogene expression, DNA repair efficiency, and sensitivity to genotoxic agents or targeted therapies. This model helps elucidate how BAP1 loss collaborates with viral oncogenes to promote cervical cancer progression, making it a powerful tool for preclinical studies.
Researchers can utilize these knockout cells for Western blotting, immunofluorescence, and flow cytometry to verify BAP1 loss and downstream effects. Functional assays such as apoptosis and proliferation measurements probe tumor suppressor activity, while co-immunoprecipitation and ChIP-qPCR dissect protein interactions and chromatin changes. RNA-seq reveals transcriptomic alterations, and drug sensitivity assays identify therapeutic vulnerabilities. For further details, please contact Ascent Research.
