The ACSL4 Knockout Ca Ski Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the Ca Ski cervical carcinoma line, designed to disrupt the ACSL4 gene. This heterogeneous knockout model enables loss-of-function studies of long-chain fatty acid-CoA ligase 4 without the need for clonal isolation, providing a versatile tool for investigating ACSL4-mediated lipid metabolism and ferroptosis regulation.
The Ca Ski cell line, isolated from a cervical epidermoid carcinoma metastasis in a Caucasian female, stably harbors the HPV-16 genome. These squamous cell carcinoma cells are a well-established model for HPV-driven oncogenesis, exhibiting dysregulated proliferation and survival pathways that intersect with metabolic reprogramming associated with cervical cancer.
ACSL4 catalyzes the conversion of polyunsaturated fatty acids such as arachidonic acid into acyl-CoA esters, a prerequisite for their incorporation into membrane phospholipids. This activity is regulated by transcription factors SREBP1 and PPAR?? and can be induced by ferroptosis inducers erastin and RSL3. ACSL4-generated PUFA-phospholipids are oxidized by ALOX15, leading to lipid peroxide accumulation and inhibition of the phospholipid hydroperoxidase GPX4, thereby executing ferroptotic cell death. Key interacting proteins include LPCAT3, TFR1, and acyl-CoA binding proteins, which facilitate lipid remodeling and iron-dependent peroxidation.
In Ca Ski cells, ACSL4 knockout enables dissection of the interplay between HPV?16-driven malignancy and ferroptosis. Given the prevalence of altered lipid metabolism in cervical cancer, this model allows investigation of whether ACSL4-dependent lipid peroxidation pathways contribute to tumor survival or drug resistance, and whether targeting ferroptosis may enhance therapeutic responses in HPV?positive carcinomas.
These polyclonal knockout cells are ideal for ferroptosis sensitivity assays using erastin/RSL3 and lipid peroxidation detection by C11?BODIPY flow cytometry or LC?MS phospholipid profiling. ACSL4 disruption can be confirmed via Western blotting and RT?qPCR, while downstream effects on GPX4 and ALOX15 expression are readily assessed. Additional applications include co?immunoprecipitation for protein interaction studies, iron assays, and phenotypic analyses such as colony formation, migration/invasion, and ATP?based viability tests. These tools support research into ferroptosis mechanisms, cervical cancer lipid metabolism, drug resistance, and HPV?related oncogenesis. For further details, contact Ascent Research.
