The CASP4 Knockout HeLa Polyclonal Cells provide a CRISPR/Cas9-mediated loss-of-function model in HeLa cells, featuring targeted disruption of the CASP4 gene. This polyclonal knockout cell population offers a heterogeneous edited pool, enabling robust assessment of CASP4-dependent processes while maintaining the inherent biological diversity of the host line. The product is intended for researchers exploring inflammatory caspase biology without the biases introduced by clonal selection.
HeLa cells are a human epithelial cervical adenocarcinoma line positive for human papillomavirus 18 (HPV-18). Widely adopted in cancer research, this immortalized line provides a well-characterized, genetically tractable system with extensive molecular annotation. Its transformed phenotype and intact cell death machinery make it an ideal background for studying pyroptosis and innate immune signaling.
CASP4 is an inflammatory caspase that senses cytosolic lipopolysaccharide (LPS), leading to its oligomerization and autoactivation. Once active, CASP4 cleaves gasdermin D (GSDMD), generating an N-terminal fragment that forms membrane pores, thereby executing pyroptosis and enabling release of IL-1?? and IL-18. Transcription of CASP4 is induced by interferon-gamma (IFN-??) via IRF1 and NF-??B, often downstream of Toll-like receptor 4 (TLR4) stimulation. The protein interacts with ASC/PYCARD, caspase-1, NLRP3, and RIPK1, positioning it at a critical junction between LPS sensing, inflammasome assembly, and inflammatory cell death.
In HeLa cells, which express the pyroptotic machinery, CASP4 knockout abolishes responses to intracellular LPS and impairs non-canonical inflammasome activation. This deficiency makes the model particularly valuable for dissecting the role of CASP4 in diseases such as sepsis, inflammatory bowel disease, colorectal cancer, and atherosclerosis. Moreover, the HPV-18 positive background permits exploration of how viral oncoproteins may intersect with CASP4-driven inflammation and cell death pathways.
This polyclonal knockout population is suited for examining non-canonical inflammasome function, pyroptosis triggered by infection or sterile stimuli, and inhibitor screening against inflammatory caspases. Common downstream assays include LDH release and propidium iodide uptake to measure cell death, ELISA for IL-1?? and IL-18, Western blotting for CASP4 and GSDMD cleavage, caspase activity assays, and immunofluorescence to visualize ASC specks and GSDMD localization. Co-immunoprecipitation can further map CASP4 interaction networks. Thus, the model supports mechanistic studies and translational research in inflammation and oncology. For further information, contact Ascent Research.
