The BAD Knockout A2780 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population derived from the human ovarian carcinoma A2780 cell line, designed to disrupt the endogenous BAD gene and create a loss-of-function model for investigating apoptosis regulation. This heterogeneous knockout pool serves as a versatile tool for dissecting BAD-mediated signaling events without the constraints of clonal selection, enabling robust characterization of BH3-only protein biology in a disease-relevant epithelial context.
The host A2780 cell line was originally established from an untreated ovarian adenocarcinoma patient and exhibits characteristic epithelial morphology. Widely employed as a model system in ovarian cancer research, A2780 cells retain key molecular features of the disease, including functional p53 and sensitivity to platinum-based chemotherapeutics, making them particularly informative for apoptosis and drug resistance studies.
BAD (BCL2-associated agonist of cell death) is a BH3-only pro-apoptotic member of the BCL-2 family that promotes mitochondrial outer membrane permeabilization (MOMP) by heterodimerizing with anti-apoptotic guardians BCL-2, BCL-XL, and BCL-W, thereby liberating BAX and BAK to oligomerize, release cytochrome c, and activate the caspase-9/caspase-3 cascade. BAD??s apoptotic function is tightly regulated by survival kinases including AKT, MAPK/ERK, PKA, and RSK, which phosphorylate BAD at Ser112, Ser136, and Ser155 to create 14-3-3 binding sites, resulting in cytoplasmic sequestration and inhibition of its mitochondrial translocation. Additionally, BAD interacts with protein phosphatases PP2A and PP1 and calcineurin, which can reverse its inactivation.
In ovarian carcinoma, BAD is often inactivated through hyperphosphorylation driven by aberrant PI3K/AKT and MAPK pathway activation, contributing to evasion of apoptosis and resistance to chemotherapy. The A2780 cell line, which displays an epithelial phenotype and intrinsic sensitivity to DNA-damaging agents, provides an ideal background to interrogate how BAD disruption alters the balance between pro- and anti-apoptotic BCL-2 family members and influences cellular responses to standard-of-care drugs such as cisplatin and paclitaxel.
Researchers can employ this polyclonal knockout model to study apoptosis signaling by co-immunoprecipitating BCL-2 family complexes, performing phospho-specific Western blotting for BAD (Ser112/136/155) and downstream caspases, and measuring mitochondrial depolarization with JC-1. Flow cytometry?Cbased Annexin V/PI staining and caspase-3/7 activity assays enable quantitative assessment of cell death induction, while viability assays (MTT) and drug sensitivity profiling across a panel of chemotherapeutics or kinase inhibitors uncover BAD-dependent resistance mechanisms. Further applications include BH3 mimetic evaluation and investigations of EGFR and JAK/STAT crosstalk. For further details or technical support, please contact Ascent Research.
