The CASP4 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population generated from the HT29 human colorectal adenocarcinoma cell line (Homo sapiens). This heterogeneous cell pool harbors targeted disruption of the CASP4 gene, avoiding clonal selection to provide a population-level loss-of-function model. It is designed for investigating CASP4-dependent signaling and pyroptotic pathways in an intestinal epithelial context.
The HT29 cell line, derived from a 44-year-old female colorectal adenocarcinoma, carries well-characterized mutations in APC and TP53 and serves as a widely used model of colorectal cancer. Its epithelial origin and retained innate immune signaling components make it particularly suitable for examining CASP4 function in mucosal inflammation, host-microbe interactions, and oncogenic processes.
CASP4 is an inflammatory caspase that acts as an intracellular sensor for lipopolysaccharide (LPS). Upon cytosolic LPS binding, CASP4 oligomerizes and cleaves gasdermin D (GSDMD), forming membrane pores that drive pyroptotic lysis and release of IL-1?? and HMGB1. This non-canonical inflammasome activation can subsequently engage the NLRP3 inflammasome and caspase-1, amplifying inflammatory responses. CASP4 expression is regulated upstream by TLR4/LPS, IFN-??/STAT1, TNF-??/NF-??B, and ER stress (PERK/CHOP) pathways. The protein directly interacts with LPS and functionally cooperates with GSDMD, ASC, NLRP3, and caspase-1.
In the HT29 colorectal cancer background, CASP4 knockout enables precise dissection of inflammatory caspase roles in tumor-promoting inflammation and epithelial innate immune responses. This model is particularly valuable for studying the contributions of non-canonical inflammasome signaling to intestinal barrier dysfunction, inflammatory bowel disease pathology, and sepsis-related epithelial injury. The polyclonal format preserves population-level responses, avoiding clonal artifacts and reflecting the cellular heterogeneity typical of tumor microenvironments.
This knockout tool supports a wide range of research applications, including inflammasome biology, innate immunity, sepsis modeling, and anti-inflammatory drug screening. Experimental readouts commonly rely on Western blotting for CASP4 and GSDMD cleavage, LDH release assays for pyroptosis, IL-1?? ELISA for cytokine maturation, and confocal microscopy for GSDMD pore formation. Co-immunoprecipitation can assess direct LPS-CASP4 interactions, while RT-qPCR and RNA-seq provide transcriptional insights. For technical inquiries or to discuss custom experimental designs, please contact Ascent Research.
