ACSL4 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human near-haploid cell line. This product provides a loss-of-function model for the ACSL4 gene, generated by CRISPR/Cas9-mediated gene disruption. The polyclonal nature reflects a mixed population of edited cells, offering a robust tool for studying ACSL4-dependent processes without clonal selection biases. Designed for functional genomics, this knockout model enables investigation of ACSL4’s role in lipid metabolism and ferroptosis regulation.
The HAP1 cell line is a male near-haploid line derived from the KBM-7 chronic myeloid leukemia cells. Its near-haploid karyotype simplifies genetic manipulation and phenotypic analysis, making it an ideal host for CRISPR-based knockout studies. HAP1 cells retain key signaling pathways relevant to cancer and metabolic research, providing a versatile platform for dissecting gene function in a leukemic cell context. The cell line’s haploid state facilitates efficient gene targeting and subsequent functional characterization.
ACSL4 encodes acyl-CoA synthetase long-chain family member 4, which converts long-chain polyunsaturated fatty acids into acyl-CoA esters, critically enriching membrane phospholipids with oxidizable acyl chains. ACSL4 is a key regulator of ferroptosis, interacting with GPX4 as a functional antagonist and cooperating with LPCAT3 to incorporate arachidonic acid into phosphatidylethanolamine. Its expression is regulated by upstream factors such as SREBP1, PPAR??, and TFEB, and repressed by TP53. ACSL4 activity promotes lipid peroxidation downstream of ferroptosis inducers like RSL3 and erastin, feeding into pathways involving ALOX5 and ALOX12. The gene also interfaces with the system xc- transporter SLC7A11 and the iron-import receptor TFRC, linking amino acid and iron metabolism to ferroptosis execution.
In the HAP1 background, loss of ACSL4 disrupts the conversion of free fatty acids to acyl-CoA esters, reducing the pool of peroxidation-susceptible phospholipids. This renders the cells resistant to ferroptotic death triggered by pharmacological agents such as RSL3 and erastin, or by genetic depletion of GPX4. The polyclonal knockout population provides a consistent model to study ferroptosis resistance mechanisms and to identify compensatory pathways. Because HAP1 cells sustain key signaling networks, the ACSL4 knockout cells allow interrogation of crosstalk between ferroptosis and other cell death modalities in a relevant leukemic context, contributing to understanding drug-resistant cancer phenotypes.
Researchers can employ these polyclonal ACSL4-knockout HAP1 cells to investigate ferroptosis mechanisms using lipid peroxidation probes such as C11-BODIPY, to validate hits from genome-wide CRISPR screens, and to perform drug screening for ferroptosis modulators. The model supports assays including Western blotting for ACSL4 and GPX4, RT-qPCR for ACSL4 and SLC7A11, cell viability assays (MTT, ATP), and phospholipidomic analysis by LC-MS. Applications extend to ischemia-reperfusion injury modeling and studies of lipid metabolism in neurodegeneration. For further details and technical support, please contact Ascent Research.
