The BAD Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the BAD gene in the human tongue squamous cell carcinoma line CAL-27. This polyclonal product provides a heterogeneous loss-of-function model with disrupted BAD expression, enabling studies of apoptosis signaling without clonal bias.
CAL-27 is an adherent epithelial cell line derived from a tongue squamous cell carcinoma and serves as a standard in vitro model for oral cancer research. It exhibits characteristic genetic alterations, including p53 mutation and aberrant PI3K/AKT pathway activation, making it a relevant platform for investigating apoptosis defects and therapeutic resistance in head and neck malignancies.
BAD encodes a pro-apoptotic BH3-only protein that senses growth factor deprivation and promotes cell death by neutralizing anti-apoptotic BCL-2 and BCL-XL. Under survival signaling, AKT, PKA, and RAS/MAPK kinases phosphorylate BAD at Ser112, Ser136, and Ser155, promoting 14-3-3 binding and cytoplasmic retention. When dephosphorylated, BAD translocates to mitochondria, heterodimerizes with BCL-2/BCL-XL, and releases BAK and BAX to permeabilize the outer mitochondrial membrane, leading to cytochrome c release, caspase-9 activation, and caspase-3 cleavage. BAD integrates upstream signals from receptors such as EGFR and IGF-1R to regulate the intrinsic apoptotic pathway.
In CAL-27 cells, hyperactivation of survival kinases, particularly AKT, results in constitutive BAD phosphorylation and apoptosis evasion. Ablation of BAD in this polyclonal knockout model allows dissection of the dependence on BAD-mediated death signaling and facilitates exploration of alternative apoptotic pathways. This model is especially applicable to studies of drug-induced apoptosis in oral cancer, including responses to cisplatin, cetuximab, and BH3 mimetics, where BAD-independent mechanisms may be targeted.
This knockout model supports a range of assays, including western blotting for total and phospho-BAD, annexin V/propidium iodide flow cytometry, caspase-3/7 activity measurements, JC-1 mitochondrial membrane potential analysis, and cell viability assays (MTT, resazurin). Co-immunoprecipitation can confirm loss of BAD-BCL-2 complexes, and scratch wound migration assays may reveal apoptosis-linked migratory changes. Key applications include investigating apoptosis resistance in head and neck squamous cell carcinoma, screening for pro-apoptotic drugs that bypass BAD, and assessing BAD as a biomarker for chemotherapy sensitivity. For further details, contact Ascent Research.
