PLA2G4A Knockout Cell Line (A549)

The choice depends on whether you are studying PLA2G4A (cytosolic phospholipase A2α, cPLA2α)'s role as the principal arachidonic acid-releasing enzyme upstream of eicosanoid biosynthesis or its functions in inflammation and respiratory disease. The Knockout line is the standard tool for asking whether cPLA2α is required for stimulus-induced arachidonic acid release — cPLA2α hydrolyzes sn-2 arachidonic acid from membrane phospholipids in response to calcium influx and MAPK phosphorylation (S505), providing substrate for COX/LOX-mediated prostaglandin and leukotriene synthesis. Overexpression is useful for studying cPLA2α in eicosanoid biosynthesis contexts. For respiratory inflammation research, the EDITGENE PLA2G4A Knockout in A-549 is highly relevant — A-549 is a lung epithelial cancer model, and cPLA2α is central to airway inflammation and asthma pathophysiology. Rescue with wild-type, lipase-dead (S228A), or phosphorylation-resistant (S505A) cPLA2α enables comprehensive structure-function studies. The knockout is a critical specificity control for cPLA2α-selective inhibitors (giripladib, ecopladib) in asthma and inflammation drug development.

Primary applications: • Arachidonic acid release: ³H-arachidonate release from prelabeled phospholipids following A23187 or PMA stimulation to quantify cPLA2α activity. • Eicosanoid generation: PGE2, LTB4, and TXB2 quantification by ELISA or mass spectrometry to characterize downstream effects of cPLA2α loss. • Calcium- and phospho-dependence: rescue with S228A (lipase-dead) or S505A (phospho-deficient) cPLA2α for structure-function dissection. • Asthma/inflammation pharmacology: critical genetic control for giripladib, ecopladib, and other cPLA2α inhibitors in respiratory disease drug development. EDITGENE recommends this model for researchers investigating arachidonic acid release, eicosanoid biosynthesis, and cPLA2α-targeted respiratory therapeutic development.

Yes. cPLA2α rescue experiments are well-established for inflammation research: • Construct design: use a codon-modified PLA2G4A sequence with a small C-terminal tag (FLAG, HA). cPLA2α has C2 calcium-binding domain and catalytic domain — preserve all elements. • Lipase-dead rescue: the S228A mutation in the catalytic serine abolishes phospholipase activity and is the standard specificity control. • Phosphorylation-deficient rescue: the S505A mutation eliminates MAPK/ERK-mediated activation phosphorylation, enabling studies of regulatory versus constitutive activity. • Functional readout: rescue should restore arachidonic acid release and downstream eicosanoid generation. A-549 transduces efficiently with lentivirus and supports stable rescue line generation.