The ACSL4 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited population in which the ACSL4 gene has been disrupted, generating a functional knockout model for studying long-chain acyl-CoA synthetase 4. This polyclonal knockout product provides a heterogeneous loss-of-function system ideal for investigating ACSL4-dependent processes without the limitations of clonal variability.
The host cell line HeLa is a human cervical adenocarcinoma-derived immortalized epithelial cell line, widely utilized as a model in cancer biology, cell signaling, and drug susceptibility studies due to its robust growth characteristics and extensive characterization.
ACSL4 catalyzes the conversion of long-chain fatty acids, particularly arachidonic acid and eicosapentaenoic acid, into fatty acyl-CoA esters, which serves as a critical step in lipid metabolism. ACSL4 preferentially esterifies polyunsaturated fatty acids (PUFAs), and these PUFA-CoAs are then incorporated into membrane phospholipids by lysophosphatidylcholine acyltransferase 3 (LPCAT3). This enrichment of PUFA-containing phosphatidylcholines provides substrates for lipoxygenases such as ALOX15, leading to the generation of lipid hydroperoxides??key executioners of ferroptosis. ACSL4 expression is regulated by upstream transcription factors including SREBP1, PPAR??, and NRF2, while its activity is functionally intertwined with GPX4, which reduces lipid hydroperoxides, and with voltage-dependent anion channels VDAC2/3. The ACSL4-LPCAT3-ALOX15 axis thus forms a pivotal node in ferroptosis signaling, controlling the abundance of oxidizable phospholipids that determine susceptibility to this iron-dependent cell death pathway.
In the HeLa context, ACSL4 disruption eliminates the primary route for PUFA esterification into membrane phospholipids, resulting in profound resistance to ferroptotic stimuli such as erastin or RSL3. This polyclonal knockout model enables researchers to dissect ferroptosis-related pathways in a well-characterized cervical cancer background, providing insights into how lipid peroxidation contributes to tumor cell survival and drug resistance.
The ACSL4 Knockout HeLa Polyclonal Cells are designed for diverse applications including mechanistic studies of ferroptosis execution, exploration of cancer cell resistance to chemotherapeutics that induce lipid peroxidation, and screening for novel ferroptosis modulators. These cells are also valuable for investigating lipid metabolism reprogramming in tumor contexts and for metabolic disease modeling where ACSL4-dependent phospholipid remodeling is implicated. Compatible assays include lipid peroxidation measurements using C11-BODIPY or MDA detection, cell viability assessments following treatment with ferroptosis inducers like erastin, RSL3, or FIN56, western blotting and RT-qPCR for ACSL4 and GPX4 expression analysis, immunofluorescence microscopy for protein localization, co-immunoprecipitation with GPX4, and phospholipidomic profiling. For further information, please contact Ascent Research.
