The CASP9 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the 786-O renal cell adenocarcinoma line, featuring targeted disruption of the CASP9 gene. This model provides a loss-of-function system for studying the initiator caspase of intrinsic apoptosis without clonal selection, enabling robust investigation of apoptotic signaling in a kidney cancer background.
The 786-O line is a clear cell renal cell carcinoma model harboring a VHL mutation, leading to constitutive HIF activation and uncontrolled proliferation. As a widely used human kidney cancer model, it offers a disease-relevant platform for evaluating the impact of CASP9 loss on apoptosis, drug sensitivity, and tumorigenesis.
CASP9 is activated upon cytochrome c (CYCS) binding to APAF1, forming the apoptosome, after mitochondrial outer membrane permeabilization driven by BAX/BAK. Active CASP9 cleaves executioner caspases-3 and -7, leading to PARP1 degradation. Upstream regulators include anti-apoptotic BCL2 and HSP70, while XIAP and BIRC5 directly inhibit CASP9. SMAC/DIABLO promotes apoptosis by neutralizing XIAP. Thus, CASP9 integrates mitochondrial damage signals to execute cell death.
In 786-O cells, CASP9 knockout disrupts intrinsic apoptosis, enabling studies of VHL-mutant renal cancer resistance to therapies. This polyclonal population is useful for examining how CASP9 loss affects drug sensitivity, hypoxia responses, and apoptotic signaling pathways, potentially revealing synthetic lethal targets or biomarker candidates in kidney cancer.
Key assays include Western blotting for cleaved CASP3/CASP7 and PARP1, caspase activity measurements, Annexin V/PI flow cytometry, cytochrome c release assays, and viability screens with MTT or CellTiter-Glo. Co-immunoprecipitation can assess interactions with APAF1 or XIAP. The model supports screening of pro-apoptotic compounds and studies of apoptosis deficiency. For further information, contact Ascent Research.
