The GSDMD Knockout DLD-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human DLD-1 colorectal adenocarcinoma cell line. This product features targeted disruption of the GSDMD gene, creating a loss-of-function model for studying pyroptosis without requiring clonal isolation. The polyclonal format retains cellular heterogeneity, enabling robust assessment of GSDMD-dependent processes in an epithelial cancer background.
The DLD-1 host cell line is an adherent epithelial line established from a Duke??s type C colorectal adenocarcinoma. It is widely used in cancer biology for investigating oncogenic signaling, drug responses, and inflammatory pathways. As DLD-1 cells express components of inflammasome machinery, they provide a relevant context for interrogating GSDMD-mediated pyroptosis in colorectal cancer and related inflammatory disorders.
GSDMD acts as the executioner of pyroptosis, a lytic, pro-inflammatory cell death. Inflammasome-activated caspases, such as caspase-1 and caspase-4/5/11, cleave GSDMD at Asp275, releasing the N-terminal pore-forming domain. This fragment oligomerizes and inserts into the plasma membrane, forming pores that induce cell swelling and rupture, and facilitate the release of mature IL-1?? and IL-18. Upstream regulators include NLRP3, NLRC4, and AIM2 inflammasomes, which recruit ASC and caspase-1, while non-canonical activation involves caspase-4/5/11 sensing cytosolic lipopolysaccharide. GSDMD also interacts with NEK7, a direct NLRP3 interactor, and its cleavage is influenced by type I interferons.
In DLD-1 colorectal adenocarcinoma cells, GSDMD knockout eliminates the primary pore-forming effector, enabling investigation of pyroptosis in cancer-associated inflammation. This loss-of-function model is suited for studying inflammasome signaling, cytokine release, and the interplay between pyroptosis and tumor cell death. Because DLD-1 cells harbor key oncogenic mutations, this model also facilitates examination of GSDMD??s role in disease contexts such as inflammatory bowel disease and sepsis.
Typical applications include western blotting for GSDMD cleavage, LDH release assays to quantify pyroptotic death, and IL-1?? ELISA to measure cytokine release. Immunofluorescence imaging and flow cytometry for propidium iodide uptake can assess pore formation and membrane integrity. These cells are amenable to high-throughput screening of pyroptosis inhibitors and mechanistic studies of inflammasome regulation using NLRP3 activators or LPS transfection. For further information, please contact Ascent Research.
