The ALOX5AP Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population in which the ALOX5AP gene has been disrupted to create a loss-of-function model of 5-lipoxygenase activating protein (FLAP). This heterogeneous pool of A-549 cells retains genetic diversity while abolishing FLAP expression, providing a robust and representative platform for interrogating FLAP-dependent pathways without the constraints of clonal selection.
The host A-549 cell line is a well-characterized human lung adenocarcinoma epithelial line originally isolated from a 58-year-old Caucasian male. It displays an adherent morphology and is established as a standard model for type II alveolar epithelial cells, widely employed in lung cancer biology, respiratory disease research, and the study of pulmonary epithelial physiology.
ALOX5AP encodes FLAP, a scaffold that shuttles arachidonic acid to ALOX5 for leukotriene biosynthesis. This process is activated by cytokines (IL-4, IL-13), calcium flux, and MAP kinase signaling, and it drives production of pro-inflammatory mediators including LTB4 and cysteinyl leukotrienes via cPLA2, LTC4 synthase, and MAPKAPK2 interactions. Downstream signaling through BLT1, BLT2, CysLT1, and CysLT2 receptors amplifies inflammatory responses, linking FLAP to diseases such as asthma, atherosclerosis, and myocardial infarction.
In the A-549 lung adenocarcinoma background, ALOX5AP knockout disrupts leukotriene and 5-HETE biosynthesis, enabling detailed examination of how FLAP-dependent lipid mediators modulate cancer cell behaviors such as proliferation, migration, and invasion. The type II alveolar cell-like characteristics of A-549 further allow investigation of FLAP in airway epithelial inflammation, making this model suitable for asthma and acute lung injury studies. Additionally, the model supports validation of therapeutic FLAP inhibitors in a disease-relevant cellular context.
Researchers can employ these polyclonal cells for quantitative leukotriene analysis by ELISA or HPLC, Western blotting for FLAP and ALOX5, RT-qPCR gene expression profiling, and arachidonic acid metabolic assays. Functional applications include cell migration/invasion studies and drug sensitivity testing with FLAP inhibitors such as MK-886 or GSK2190915, enabling drug target validation and inflammation signaling research. For additional technical inquiries, please contact Ascent Research.
