The CASP4 Knockout Huh-7 Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population derived from the human HuH-7 hepatocellular carcinoma line, designed to abolish functional CASP4 expression. This pooled knockout product provides a heterogeneous loss-of-function model suitable for studying caspase-4-dependent biology without clonal limitations. The polyclonal format maintains genetic diversity while ensuring target-gene disruption across the population, making it ideal for robust, reproducible experiments in immunology and cancer research.
The HuH-7 cell line originated from a well-differentiated hepatocellular carcinoma resected from a 57-year-old Japanese male and is widely employed as a model for liver cell biology, metabolic functions, and hepatitis C virus replication. HuH-7 cells retain key hepatocytic features, including protein synthesis and secretion pathways, and are extensively used in hepatic inflammation, fibrosis, and drug metabolism studies. Their human origin and tumor background provide a clinically relevant platform for investigating molecular mechanisms in hepatocellular carcinoma and innate immune responses in the liver.
CASP4 encodes caspase-4, an inflammatory protease that senses intracellular lipopolysaccharide (LPS) via its CARD domain, triggering oligomerization and autoproteolytic activation. Activated caspase-4 cleaves gasdermin D (GSDMD), generating a pore-forming N-terminal fragment that induces pyroptosis and the release of IL-1??, IL-18, and DAMPs. This non-canonical inflammasome pathway is regulated by TLR4 and type I interferon signaling, which upregulate CASP4 expression through NF-??B. Caspase-4 activity also promotes NLRP3 inflammasome assembly and caspase-1-mediated cytokine maturation.
In the HuH-7 hepatic context, CASP4 disruption provides a unique tool to deconvolute non-canonical inflammasome signaling in hepatocellular carcinoma cells. Hepatocytes are constantly exposed to gut-derived microbial products and respond via conserved innate immune pathways; thus, loss of caspase-4 can elucidate its contribution to LPS-driven hepatocyte death and inflammation. This model is particularly relevant for examining how pyroptotic signaling intersects with tumor cell survival, immune evasion, and liver disease progression, including non-alcoholic steatohepatitis and endotoxemia-induced liver injury.
These polyclonal knockout cells are suited for applications ranging from mechanistic studies of pyroptosis and inflammasome regulation to drug discovery screens for caspase-4 or GSDMD inhibitors. Researchers can employ assays such as Western blotting to monitor GSDMD cleavage and caspase-4 activation, LDH release or propidium iodide uptake to quantify pyroptotic cell death, and ELISA for IL-1??/IL-18 secretion. Co-immunoprecipitation and confocal microscopy further enable analyses of protein complexes and pore formation. This model also facilitates host-pathogen interaction studies with Gram-negative bacteria and evaluation of therapeutics targeting sepsis, inflammatory bowel disease, or liver fibrosis. For further details, please contact Ascent Research.
