This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from 769-P human clear cell renal cell carcinoma cells, featuring targeted disruption of the BAD gene. The polyclonal format provides a heterogeneous loss-of-function model suitable for studying BAD-dependent signaling in a tumor-derived epithelial background. The knockout model is generated via CRISPR/Cas9-mediated gene disruption, enabling robust investigation of BAD??s role in apoptosis and survival pathways without the clonal selection constraints of monoclonal lines.
The 769-P cell line was established from a primary clear cell renal cell carcinoma and serves as a well-characterized model of kidney epithelium-derived tumors. These cells exhibit features typical of renal cancer, including dysregulated apoptotic signaling and reliance on pro-survival networks, making them an appropriate host for studying BAD-mediated apoptosis. The parental line retains sensitivity to growth factor and kinase inhibitor perturbations, providing a physiologically relevant context for dissecting intrinsic and extrinsic cell death mechanisms.
BAD is a pro-apoptotic BH3-only member of the BCL-2 family that functions at a critical regulatory node in the intrinsic apoptosis pathway. In its unphosphorylated state, BAD heterodimerizes with anti-apoptotic proteins such as BCL-2, BCL-XL, and BCL-W, liberating BAX and BAK to trigger mitochondrial outer membrane permeabilization, cytochrome c release, and subsequent activation of caspase-9 and caspase-3. Phosphorylation by AKT, MAPK, or PKA downstream of PI3K and growth factor receptors (including IGF-1R and EGFR) creates 14-3-3 binding sites, sequestering BAD in the cytosol and suppressing apoptosis. BAD also interacts with BIM, BID, and PP2A, fine-tuning the apoptotic threshold.
In renal cell carcinoma, aberrant activation of the PI3K/AKT pathway and downstream BAD phosphorylation contribute to chemoresistance and tumor maintenance. Disrupted BAD function in the 769-P knockout model eliminates a key pro-apoptotic constraint, allowing researchers to assess the dependence of this tumor type on BAD-mediated cell death suppression. This model is particularly relevant for exploring how MAPK/ERK and PI3K/AKT survival signaling converges on BAD to modulate mitochondrial integrity and therapy response. Comparative studies using the knockout cells can reveal whether BAD inactivation promotes resistance to agents that induce intrinsic apoptosis, providing insight into therapeutic vulnerabilities.
Researchers can employ this polyclonal BAD knockout population to investigate apoptosis regulation, PI3K/AKT-driven survival mechanisms, and chemosensitivity in renal cell carcinoma. Representative assays include western blotting for total and phospho-BAD, Annexin V/PI apoptosis assays, co-immunoprecipitation of BAD with BCL-2 or 14-3-3, flow cytometry for mitochondrial membrane potential, and cell viability assays with PI3K/AKT inhibitors. Caspase-3/7 activity provides direct readouts of effector caspase engagement. For further technical information, please contact Ascent Research.
