The GSDMD Knockout HT29 Polyclonal Cells product comprises a population of HT29 human colorectal adenocarcinoma cells that have been genetically modified using CRISPR/Cas9 to disrupt the GSDMD gene. This heterogeneous polyclonal population provides a loss-of-function model for studying the role of GSDMD in pyroptosis and inflammasome signaling.
The HT29 cell line is a widely used model of human intestinal epithelium, originally established from a primary colorectal adenocarcinoma. These cells retain properties of enterocytic differentiation, including the capacity for mucus production, and are commonly employed in studies of gastrointestinal biology, host-microbe interactions, and oncogenic signaling.
GSDMD functions as the pore-forming executor of pyroptosis downstream of inflammasome activation. Canonical inflammasomes (e.g., NLRP3, NLRC4, AIM2) mediate caspase-1 activation, whereas non-canonical signaling involves direct cleavage of GSDMD by caspase-4/5 in humans or caspase-11 in mice. Upon cleavage, the N-terminal fragment of GSDMD translocates to the plasma membrane and interacts with phosphatidylinositol 4-phosphate and phosphatidylserine, oligomerizes, and forms membrane pores. These pores compromise cellular osmotic integrity, resulting in cell swelling, lysis, and the passive release of mature IL-1?? and IL-18, as well as alarmins such as HMGB1 and LDH.
In the HT29 intestinal epithelial context, disruption of GSDMD expression permits the dissection of pyroptosis-dependent inflammatory responses from other programmed cell death modalities. Given the critical role of the intestinal epithelium in sensing microbial products and releasing cytokines, this knockout model is particularly valuable for investigating the contributions of GSDMD to epithelial barrier function, host defense against enteric pathogens, and the pathogenesis of inflammatory bowel diseases including Crohn??s disease and ulcerative colitis.
Researchers can utilize this polyclonal knockout cell population to investigate GSDMD-dependent mechanisms in canonical and non-canonical inflammasome pathways, to screen for pharmacological modulators of pyroptosis, or to explore the interplay between pyroptosis and tumor cell biology. Typical assays include immunoblotting for GSDMD and caspases, LDH release and propidium iodide uptake to measure cell lysis, ELISA-based quantification of IL-1?? and IL-18 secretion, and immunofluorescence imaging of pore formation. For additional details or to inquire about custom applications, please contact Ascent Research.
