The GSDMD Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered from the human osteosarcoma cell line 143B, designed to disrupt the GSDMD gene. This heterogeneous loss-of-function model enables investigation of gasdermin D-dependent pyroptosis without clonal selection bias, providing a versatile platform for downstream functional assays.
The 143B cell line, a highly metastatic human osteosarcoma derivative of TE-85 from a female patient, exhibits osteoblastic properties including matrix production and osteoid formation. Its aggressive tumorigenic behavior and osteoblast-like characteristics make it particularly suitable for studying bone cancer progression and the tumor microenvironment’s inflammatory components.
GSDMD is the pore-forming executioner of pyroptosis, a pathway activated upon inflammasome assembly. In response to stimuli, sensor proteins such as NLRP3, AIM2, NLRC4, and Pyrin recruit ASC and caspase-1, which cleaves GSDMD into its active N-terminal fragment. This fragment oligomerizes in the plasma membrane, creating pores that permit the release of IL-1?? and IL-18, as well as HMGB1 and LDH, and trigger cell swelling and lysis. Additional regulation involves non-canonical inflammasomes via caspase-4/5/11, and upstream signals from TLR ligands, TNF-??, and NF-??B. Downstream, potassium efflux and NLRP3 inflammasome amplification propagate the response. GSDMD interaction with lipids like cardiolipin and phosphatidylinositol phosphates facilitates pore formation, and its oligomers are key for ASC speck-dependent cytokine secretion.
In osteosarcoma, GSDMD-mediated pyroptosis may influence tumor cell survival, metastatic dissemination, and the immune landscape. By abrogating GSDMD function in this aggressive background, researchers can examine the protein’s contributions to chemotherapy resistance and the secretion of pro-inflammatory mediators, offering mechanistic insights into bone malignancy therapies.
This polyclonal GSDMD knockout cell population is suited for pyroptosis mechanism studies, inflammasome signaling analysis, and cancer drug testing. Representative assays include Western blotting for protein expression, LDH release and flow cytometry for cell death, IL-1??/IL-18 ELISA for cytokine quantification, immunofluorescence to visualize pore formation or ASC specks, real-time qPCR, and caspase-1 activity measurements. Applicable disease models include sepsis, inflammatory bowel disease, gout, atherosclerosis, melanoma, gastric cancer, and osteosarcoma progression. For further details, please contact Ascent Research.
