The CASP3 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Huh-7 hepatocellular carcinoma line, with targeted disruption of the CASP3 executioner caspase gene. This heterogeneous pool lacks functional caspase-3 protein, enabling studies of apoptosis resistance in a liver cancer context without clonal selection artifacts.
Huh-7 cells, originating from a well-differentiated liver tumor of a 57-year-old Japanese male, are a widely used epithelial HCC model with hepatocyte-like characteristics and sensitivity to death receptor ligands (TNF, FASL) and mitochondrial apoptotic signals. Their robust response to apoptotic stimuli makes them an optimal host for engineering a caspase-3-deficient model.
Caspase-3 is the pivotal executioner protease in the apoptosis cascade, activated downstream of initiators CASP8 (extrinsic pathway) and CASP9 (intrinsic pathway) following BAX/BAK-mediated cytochrome c release and APAF1 apoptosome assembly, or death receptor activation. It cleaves key substrates such as PARP1, DFFA/ICAD, and cytokeratins to dismantle the cell, and is regulated by XIAP and BCL2 family proteins. CASP3 knockout blocks this terminal cleavage, conferring apoptosis resistance and providing a unique tool to interrogate caspase-3-dependent signaling.
In hepatocellular carcinoma, apoptosis evasion is a critical hallmark. This knockout model allows direct investigation of how liver cancer cells survive without caspase-3, facilitating dissection of alternative death pathways (e.g., pyroptosis, necrosis) and evaluation of therapeutic vulnerabilities. It is invaluable for studying drug sensitivity and resistance mechanisms where caspase-3 activation is a central pharmacodynamic endpoint.
Researchers can employ these cells in western blotting for cleaved caspase-3/ PARP1 as apoptosis-negative controls, Annexin V/PI flow cytometry, TUNEL assays, and RT-qPCR profiling. Drug screening applications include identifying sensitizers that restore apoptosis or agents that induce caspase-3-independent lethality. This model also supports 3D culture and co-culture experiments to mimic the tumor microenvironment under impaired apoptosis. For further details, please contact Ascent Research.
