The CASP4 Knockout 786-O Polyclonal Cells product comprises a population of human 786-O renal cell adenocarcinoma cells engineered via CRISPR/Cas9-mediated gene disruption to ablate CASP4 expression. This polyclonal knockout cell population is generated without single-cell cloning, providing a heterogeneous pool of edited alleles that collectively yield loss of CASP4 function. It serves as a versatile tool for investigating caspase-4-dependent signaling mechanisms in a relevant cancer cell background. The product is designed for researchers requiring a robust loss-of-function model to dissect pathways regulated by this inflammatory caspase.
The parental 786-O cell line is a well-characterized model derived from a human clear cell renal cell carcinoma (ccRCC) and harbors a naturally occurring VHL tumor suppressor mutation, a hallmark of the majority of ccRCC cases. These cells retain epithelial morphology and are widely employed to study renal cancer biology, hypoxia signaling, and tumor microenvironment interactions. The VHL-deficient background makes the 786-O line particularly valuable for exploring connections between inflammatory signaling, metabolic reprogramming, and oncogenic processes.
CASP4 encodes caspase-4, an inflammatory caspase that serves as an intracellular LPS receptor. Upon binding cytosolic LPS, caspase-4 activates and cleaves gasdermin D (GSDMD), generating N-terminal fragments that form membrane pores, triggering pyroptotic cell death and release of IL-1?? and IL-18. Its expression is induced by LPS-TLR4 signaling and type I interferons, and it is further regulated by IRE1?? under ER stress. Caspase-4 also functionally interacts with NLRP3 and ASC, bridging non-canonical and canonical inflammasome pathways.
In the context of 786-O ccRCC cells, CASP4 disruption allows for dissection of pyroptotic and inflammatory responses that may influence tumor growth, immune evasion, and therapeutic sensitivity. Renal cell carcinomas often exhibit a highly inflammatory microenvironment, and caspase-4-driven pyroptosis could contribute to the release of danger signals and cytokines that shape antitumor immunity or promote tumor progression. The availability of these CASP4 knockout 786-O polyclonal cells thus enables investigation of how loss of caspase-4 alters cytokine profiles, cell death modalities, and downstream signaling in a kidney cancer background.
Researchers can employ these knockout cells in a variety of assays to elucidate caspase-4 biology. Typical applications include monitoring GSDMD cleavage and IL-1??/IL-18 secretion by Western blotting and ELISA, quantifying cell death via LDH release or flow cytometry, and assessing transcriptional changes by RT-qPCR. Co-immunoprecipitation studies can probe caspase-4 interactions with LPS and other binding partners, while immunofluorescence imaging can visualize GSDMD pore formation. These cells are also suitable for drug screening campaigns targeting pyroptosis or inflammatory pathways. For more information, please contact Ascent Research.
