The ALOX12 Knockout NCI-H1299 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted disruption of the ALOX12 gene in the human non-small cell lung carcinoma (NSCLC) cell line NCI-H1299. This loss-of-function model provides a versatile platform for studying arachidonate 12-lipoxygenase biology in lung cancer, ferroptosis, and eicosanoid signaling.
NCI-H1299 is an adherent epithelial cell line derived from a lymph node metastasis of a lung carcinoma. These p53-null cells are widely employed as a model for NSCLC tumorigenesis, metastatic progression, and drug sensitivity, particularly in p53-deficient tumor contexts.
ALOX12 encodes a 12-lipoxygenase that catalyzes the conversion of arachidonic acid to 12-hydroperoxyeicosatetraenoic acid (12-HPETE), a key mediator of lipid peroxidation and ferroptosis. The enzyme is transcriptionally regulated by p53 (TP53), nuclear factor erythroid 2-related factor 2 (NRF2), and inflammatory cytokines such as tumor necrosis factor-alpha (TNF-??). Its lipid product 12-HETE signals via G protein-coupled receptor 31 (GPR31), promoting reactive oxygen species (ROS) generation and mitogen-activated protein kinase (MAPK) pathway activation. ALOX12 functions within the p53/ALOX12/12-HETE/GPX4 ferroptosis axis, where GPX4 counteracts ALOX12-driven lipid peroxidation. The enzyme also interacts with phospholipase A2 (PLA2) to release arachidonic acid from membrane phospholipids, linking it to PI3K/AKT/mTOR and oncogenic signaling networks.
In the p53-null NCI-H1299 background, ALOX12 disruption enables examination of p53-independent ferroptosis regulation and its impact on NSCLC cell behavior. The knockout may impair ferroptosis execution, alter MAPK and PI3K/AKT signaling, and modulate sensitivity to chemotherapeutics (e.g., cisplatin) and ferroptosis inducers (e.g., erastin, RSL3). The polyclonal format retains cellular heterogeneity, reducing clonal bias and supporting robust functional analyses.
This knockout model is applicable to ferroptosis mechanism studies, lipid peroxidation assays (C11-BODIPY), eicosanoid profiling (LC-MS), drug sensitivity screening, and cell death evaluation via flow cytometry. It also supports standard techniques such as Western blotting, RT-qPCR, Transwell migration, and colony formation. Together, these applications facilitate investigation of ALOX12 in oxidative stress, eicosanoid biology, and lung cancer. For further technical information, please contact Ascent Research.
