The AIFM2 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human oral squamous cell carcinoma (OSCC) cell line CAL-27. This product targets the AIFM2 gene (also known as FSP1), which encodes a mitochondrial flavoprotein with dual roles in caspase-independent apoptosis and ferroptosis suppression. The polyclonal format reflects a mixed population of edited cells, enabling robust loss-of-function studies without clonal selection. These cells provide a versatile platform for investigating AIFM2-dependent mechanisms in the context of oral cancer biology.
The parental CAL-27 cell line was established from a human tongue squamous cell carcinoma and is widely employed as a model for oral cancer research. CAL-27 cells exhibit an epithelial morphology and carry a TP53 mutation, which alters canonical p53-mediated responses. This genetic background makes the line particularly relevant for studying alternative cell death pathways and the interplay between ferroptosis and apoptosis in TP53-mutant cancers. The cells retain key characteristics of the tumor of origin, offering a physiologically meaningful system for evaluating gene function.
AIFM2 functions downstream of TP53 to mediate caspase-independent apoptosis, characterized by chromatin condensation and DNA fragmentation. As the ferroptosis suppressor protein FSP1, it reduces coenzyme Q10 (CoQ10) to ubiquinol at the plasma membrane, which traps lipid peroxyl radicals and prevents lipid peroxidation, thereby inhibiting ferroptosis. AIFM2 is regulated by DNA damage, oxidative stress, and NFE2L2 (NRF2), and it interacts with ubiquinone, NAD(P)H, and flavin adenine dinucleotide (FAD). Within the ferroptosis pathway, AIFM2 opposes the action of GPX4 and ACSL4, while the TP53/BAX/BCL2 axis connects it to apoptosis. These molecular interactions position AIFM2 at a critical node between cell death modalities.
In the CAL-27 TP53-mutant background, AIFM2??s apoptotic function may be attenuated, whereas its role as a ferroptosis suppressor becomes particularly important for understanding drug resistance and tumor survival. Disruption of AIFM2 in this model allows researchers to dissect p53-independent ferroptosis mechanisms and evaluate the contribution of FSP1 to lipid peroxidation control in oral squamous cell carcinoma. This system is valuable for exploring how cancer cells evade ferroptosis under therapeutic stress and for identifying synthetic lethal interactions with GPX4 inhibitors or system xc- blockers.
Typical applications include assessing caspase-independent apoptosis via Annexin V/PI flow cytometry and monitoring chromatin condensation with DAPI staining. Ferroptosis susceptibility can be measured using C11-BODIPY lipid peroxidation assays, while AIFM2 expression and translocation are evaluated by western blotting and immunofluorescence. RT-qPCR analyses of p53 target genes and co-immunoprecipitation studies of AIFM2 with ubiquinone or other interactors further elaborate its signaling network. This polyclonal knockout population serves as a foundational tool for cancer biologists and pharmacologists investigating cell death regulation in oral oncology. For further details, please contact Ascent Research.
