The CASP4 Knockout NCI-H1299 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the CASP4 gene in the NCI-H1299 human lung adenocarcinoma epithelial cell line. This loss-of-function model enables investigation of CASP4-mediated biological processes without altering the native p53-null genetic background of the host cells. The polyclonal format offers a heterogeneous pool of edited alleles, facilitating population-level studies of gene function and signaling.
The NCI-H1299 cell line was originally derived from a lymph node metastasis of a human lung adenocarcinoma and is widely employed as a model for non-small cell lung cancer (NSCLC) metastasis. Due to its TP53 null status, NCI-H1299 provides a genetically defined background for examining tumor-suppressor-independent mechanisms of cancer progression and innate immune signaling. Researchers rely on this line to dissect molecular pathways governing lung adenocarcinoma metastasis and to evaluate therapeutic targets in a clinically relevant context.
CASP4 (caspase-4) is an inflammatory caspase that functions as an intracellular sensor for lipopolysaccharide (LPS) from Gram-negative bacteria. Upon direct binding to cytosolic LPS, CASP4 oligomerizes and becomes activated, leading to cleavage of gasdermin D (GSDMD). GSDMD N-terminal fragments then form pores in the plasma membrane, triggering pyroptosis??a lytic form of cell death??and facilitating the release of mature interleukin-1?? (IL-1??) and interleukin-18 (IL-18). CASP4 expression is transcriptionally regulated by type I and II interferons, IRF1, and NF-??B, linking microbial detection to inflammatory gene induction. Additionally, CASP4 intersects with the canonical NLRP3 inflammasome pathway through GSDMD-mediated membrane damage and potassium efflux.
In the context of NCI-H1299 lung adenocarcinoma cells, CASP4 knockout provides a unique platform to dissect the role of non-canonical inflammasome activation in tumor biology. Because lung epithelial cells are frequently exposed to microbial products, CASP4-mediated pyroptosis may influence the tumor microenvironment, immune cell recruitment, and cancer cell survival during infection or inflammation. The knockout model allows direct assessment of how CASP4 loss affects inflammatory cytokine secretion, cell death pathways, and potentially metastatic behavior, contributing to a deeper understanding of the interplay between innate immunity and lung cancer progression.
Researchers can employ these polyclonal knockout cells in a variety of assays to study pyroptosis and inflammasome signaling, including LPS-induced cell death measurement by LDH release or propidium iodide uptake, western blotting for CASP4 and cleaved GSDMD, ELISA for IL-1?? and IL-18, RT-qPCR analysis of inflammatory cytokine transcripts, and flow cytometry-based detection of pyroptotic cells. This product is suitable for screening modulators of the non-canonical inflammasome pathway, evaluating crosstalk between pyroptosis and apoptosis, and exploring cancer immunotherapy strategies that target innate immune sensors. For further information or custom gene-editing services, please contact Ascent Research.
