The ACSL4 Knockout NCI-H1299 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population generated from the NCI-H1299 human non-small cell lung cancer cell line. This product delivers a heterogeneous pool of cells carrying targeted disruptions in the ACSL4 gene, enabling robust loss-of-function studies without the constraints of clonal selection. The polyclonal format maintains genetic diversity while eliminating ACSL4 protein function, offering a versatile system for investigating ACSL4-mediated processes.
NCI-H1299 was established from a lymph node metastasis of a lung adenocarcinoma and serves as a classic p53-deficient NSCLC model. It is characterized by rapid proliferation, epithelial morphology, and widespread use in studies of tumor metabolism, invasion, and therapeutic resistance. Its metastatic origin and molecular profile make it particularly relevant for examining lipid-dependent cell death pathways in lung cancer.
The ACSL4 gene product catalyzes the activation of long-chain polyunsaturated fatty acids, with a strong preference for arachidonic acid, to their acyl-CoA thioesters. This critical step provides substrates for LPCAT3-mediated incorporation into membrane phospholipids. Subsequent enzymatic oxidation by lipoxygenases ALOX12 and ALOX15 generates lipid hydroperoxides that propagate membrane damage. ACSL4 expression is under the control of transcription factors SP1, SREBP1, and PPAR??, and is post-translationally regulated by nutrient-sensing pathways including mTORC1 and AMPK. Rising lipid peroxides ultimately overwhelm the GPX4-dependent detoxification system, executing ferroptotic cell death. Additionally, ACSL4 physically associates with the molecular chaperone HSP90, implicating chaperone-assisted folding or stabilization in its function.
In the NCI-H1299 background, CRISPR-mediated ACSL4 disruption attenuates arachidonoyl-CoA synthesis, thereby limiting the pool of oxidizable membrane phospholipids and conferring resistance to ferroptosis inducers such as erastin and RSL3. This phenotype allows researchers to dissect ACSL4-dependent lipid remodeling and ferroptosis signaling in a lung adenocarcinoma setting. Moreover, given the emerging role of ferroptosis in tumor suppression and drug response, this knockout model serves as a valuable platform to evaluate ACSL4 as a potential therapeutic vulnerability in NSCLC.
Typical research applications encompass ferroptosis mechanism elucidation, lipidomic profiling of phospholipid subspecies, and interrogation of ACSL4-dependent drug sensitivity. Standard experimental approaches include Western blotting for ACSL4 protein levels, RT-qPCR for transcript quantification, C11-BODIPY fluorometric lipid peroxidation assays, and cell viability analysis following erastin or RSL3 challenge. Co-immunoprecipitation can map protein?Cprotein interactions with partners such as LPCAT3 and HSP90. For additional product information or custom requests, please contact Ascent Research.
