The ALOX12 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited human lung adenocarcinoma epithelial cell population with targeted disruption of the ALOX12 gene. This polyclonal knockout pool provides a heterogeneous loss-of-function model, eliminating expression of arachidonate 12-lipoxygenase. It offers researchers a ready-to-use system to investigate ALOX12-dependent signaling without the need for clonal isolation, preserving population-level biological variation relevant to cancer cell biology.
The parental A-549 cell line, derived from a lung adenocarcinoma of a 58-year-old Caucasian male, is a widely adopted model for alveolar epithelial biology and non-small cell lung cancer. These adherent cells retain genetic features characteristic of pulmonary adenocarcinoma, including dysregulated growth factor signaling and metastatic capacity. Culturing under standard conditions ensures reproducible genetic perturbation experiments, making the A-549 background a reliable platform for exploring lipid-mediated pathways in a disease-relevant context.
ALOX12 encodes arachidonate 12-lipoxygenase, which converts arachidonic acid to 12(S)-HETE, a bioactive lipid. Its expression is regulated by EGF, TGF-??, hypoxia, p53, and SP1. 12(S)-HETE activates the GPR31 receptor, triggering MAPK/ERK and PI3K-AKT cascades and modulating NF-??B, c-Fos, and PPAR??. It interacts with ALOX5AP, 5-lipoxygenase, cytochrome P450, and Src kinases to control proliferation, apoptosis, and adhesion.
In A-549 cells, ALOX12 knockout ablates 12(S)-HETE synthesis, disrupting GPR31-driven pro-inflammatory and pro-survival pathways. This perturbation attenuates MAPK and PI3K-AKT signaling, which are frequently hyperactivated in lung adenocarcinoma to promote tumor growth and resistance to apoptosis. The knockout model thus provides a defined genetic tool to delineate the contribution of 12-lipoxygenase to cancer cell proliferation, migration, and metastasis, and to evaluate therapeutic interventions targeting this lipid axis.
Applications include lung cancer mechanism dissection, anti-tumor drug screening, and lipid signal transduction studies. Common assays involve Western blotting and RT-qPCR for ALOX12 and target genes, 12(S)-HETE quantification by ELISA or LC-MS, MTS/MTT proliferation assays, transwell migration/invasion tests, flow cytometric analysis of apoptosis and phospho-ERK, and RNA-seq transcriptomic profiling. These polyclonal knockout cells also serve in co-culture and xenograft models to probe tumor-microenvironment interactions. For additional information, please contact Ascent Research.
