The GSDME Knockout 769-P Polyclonal Cells product consists of a population of 769-P cells subjected to CRISPR/Cas9-mediated disruption of the GSDME gene, generating a polyclonal knockout pool for loss-of-function studies. This genetically heterogeneous population lacks clonal selection, preserving biological variability while effectively ablating GSDME expression across the bulk culture. The polyclonal format is well-suited for experiments where pooled knockout responses better represent heterogeneous tumor cell behavior.
769-P cells are a widely used human clear cell renal cell carcinoma (ccRCC) epithelial line established from a primary adenocarcinoma. These cells harbor a mutation in the VHL tumor suppressor gene, leading to constitutive activation of hypoxia-inducible factor (HIF) signaling. They are tumorigenic and metastatic in nude mice, making them a valuable model for studying ccRCC pathogenesis, hypoxia-driven pathways, and therapeutic responses.
GSDME (DFNA5) functions as a key executioner of pyroptosis, an inflammatory programmed cell death. Upon activation by caspase-3 or caspase-7, GSDME is cleaved to release its N-terminal domain, which oligomerizes to form membrane pores, causing cell swelling and lysis. This process triggers the release of damage-associated molecular patterns (DAMPs) such as HMGB1, along with pro-inflammatory cytokines IL-1?? and IL-18. GSDME activity is regulated by upstream signals including p53, TNF-??, NF-??B, and interferons, and it interacts with GSDMD to modulate inflammasome-driven responses. In cancer, GSDME acts as a tumor suppressor, and its expression sensitizes cells to chemotherapy-induced pyroptosis.
In 769-P cells, GSDME knockout enables researchers to dissect the switch between pyroptotic and apoptotic cell death. Loss of GSDME abrogates pyroptosis, shifting cell death to a non-inflammatory apoptotic mode, which may impair anti-tumor immune responses. This model is particularly relevant in ccRCC, where VHL/HIF-driven alterations intersect with inflammatory signaling. By comparing wild-type and GSDME-knockout 769-P cells, investigators can explore how pyroptosis contributes to tumor immunogenicity, metastasis, and sensitivity to DNA-damaging chemotherapeutics.
Typical applications include probing pyroptosis mechanisms using LDH release and HMGB1 ELISA assays, assessing caspase-3/7 cleavage by Western blotting, and evaluating inflammatory cytokine release. The polyclonal knockout cells are also suited for drug screening, such as testing cisplatin or doxorubicin, and for co-culture experiments examining immune cell recruitment. Additionally, they facilitate investigation of GSDME crosstalk with NF-??B and inflammasome pathways. For further details or technical support, please contact Ascent Research.
