The BCL2 Knockout 786-O Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted gene disruption of BCL2 in the 786-O human clear cell renal cell carcinoma line. This polyclonal pool preserves population-level heterogeneity, circumventing clonal selection biases, and is optimally suited for studies requiring robust, reproducible loss-of-function phenotypes in apoptosis and drug response investigations. The knockout model provides a versatile platform for dissecting BCL2-dependent mechanisms in a genetic background relevant to kidney cancer.
The parental 786-O line is a human epithelial cancer cell line derived from a primary clear cell adenocarcinoma of the kidney. It is characterized by a mutation in the VHL tumor suppressor gene leading to constitutive stabilization of hypoxia-inducible factors (HIFs), which drives a pseudo-hypoxic state. This well-established renal cell carcinoma model is extensively used to explore the molecular underpinnings of kidney tumorigenesis, angiogenesis, and apoptosis resistance, making it an ideal host for BCL2 knockout.
BCL2 encodes an outer mitochondrial membrane protein that inhibits apoptosis by binding and neutralizing BAX and BAK, thereby preventing mitochondrial outer membrane permeabilization, cytochrome c release, and activation of the APAF1/caspase-9 cascade and effector caspases-3 and -7. Its expression is driven by PI3K/AKT, NF-??B, and STAT3 survival signaling. BCL2 interacts with BH3-only proteins (BIM, BAD, BID, PUMA, NOXA) and multidomain family members (BCL-XL, MCL-1), acting as a nodal integrator of intrinsic apoptotic and survival signals.
BCL2 knockout in the VHL-mutant 786-O cells ablates a key survival mechanism, rendering the polyclonal population hypersensitive to intrinsic apoptosis inducers and BH3 mimetics such as venetoclax. This model uniquely enables dissection of the interplay between HIF-driven survival pathways and mitochondrial apoptosis, offering a physiologically relevant system to study drug resistance and evaluate therapeutic strategies in renal cell carcinoma.
Primary applications include mechanistic studies of apoptosis resistance, drug sensitivity profiling, functional genomics of cell death, and screening of BH3 mimetics and novel pro-apoptotic compounds. Representative assays encompass western blotting for BCL2, cleaved caspase-3, and PARP; Annexin V/propidium iodide apoptosis assays; cell viability measurements by MTT or CellTiter-Glo; cytochrome c release detection; co-immunoprecipitation of BCL2 with BAX or BIM; BH3 profiling; and RT-qPCR analysis of BCL2 family gene expression. For further inquiries and batch-specific information, please contact Ascent Research.
