The GSDMD Knockout SK-OV-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the human SK-OV-3 ovarian adenocarcinoma cell line, engineered to disrupt the endogenous GSDMD gene. This polyclonal knockout pool preserves inherent cellular heterogeneity by avoiding single-cell clonal selection, making it a physiologically relevant loss-of-function model for investigating GSDMD-dependent pyroptosis in ovarian cancer. The gene disruption eliminates GSDMD expression, providing a clean background for mechanistic studies. It is an ideal tool for dissecting the molecular pathways regulated by GSDMD without confounding artifacts from clonal variation.
The parental SK-OV-3 cell line, established from the ascitic fluid of a patient with serous ovarian adenocarcinoma, is a widely validated epithelial model for studying ovarian cancer progression, metastasis, and chemoresistance. These cells carry mutations in TP53 and other cancer-related genes, and they are frequently used to evaluate targeted therapies and to investigate signaling networks that drive tumor aggressiveness. SK-OV-3 is particularly suited for examining peritoneal dissemination and drug resistance mechanisms, key aspects of ovarian cancer lethality. Their adherent growth and tumorigenicity in xenografts make them a robust platform for functional genomic studies.
GSDMD (gasdermin D) is the executioner of pyroptosis, a programmed lytic cell death pathway essential for innate immunity. It is activated downstream of inflammasomes such as NLRP3 and AIM2, which recruit ASC/PYCARD and pro-caspase-1 to form signaling complexes. Inflammatory caspases CASP1, CASP4, and CASP5 cleave GSDMD, releasing its N-terminal domain that oligomerizes into plasma membrane pores. These pores cause cell swelling and lysis, and facilitate the release of mature IL-1?? and IL-18. Upstream triggers include LPS, ATP, and nigericin, and membrane binding is mediated by interactions with cardiolipin and phosphatidylinositol phosphates.
In ovarian cancer, pyroptosis may influence tumor progression and chemosensitivity by modulating the inflammatory milieu. SK-OV-3 cells, derived from a chemotherapy-resistant patient, provide a relevant model to examine how GSDMD-dependent cell death intersects with pathways controlling metastasis and drug response. Disrupting GSDMD in this background allows researchers to isolate its specific contributions to IL-1??/IL-18 secretion and cell-intrinsic properties such as migration, invasion, and survival under chemotherapeutic stress. This knockout model also facilitates exploration of non-pyroptotic roles of GSDMD in cancer cell biology.
Researchers can use these knockout cells to validate GSDMD cleavage by western blotting, measure caspase-1 activity, and quantify LDH release. The model supports ELISA-based cytokine profiling, immunofluorescence analysis of pore formation, and drug sensitivity assays to assess pyroptosis-inducing compounds. Migration and invasion experiments can further define GSDMD’s role in ovarian cancer aggressiveness. Additionally, co-culture assays with immune cells can be employed to study the impact of GSDMD loss on tumor-immune interactions. For more information, please contact Ascent Research.
