AGPAT2 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma cell line, with targeted disruption of the AGPAT2 gene. AGPAT2 encodes 1-acylglycerol-3-phosphate O-acyltransferase 2, a critical enzyme that converts lysophosphatidic acid (LPA) to phosphatidic acid (PA) in the Kennedy pathway. This polyclonal population contains a heterogeneous mix of loss-of-function alleles, enabling robust functional studies of AGPAT2-dependent lipid metabolism without the biases of clonal isolation.
The A-549 cell line was originally established from a human lung carcinoma and exhibits characteristics of alveolar type II pneumocytes, making it a standard model for lung adenocarcinoma and pulmonary epithelial biology. These cells maintain high proliferative capacity and metabolic flexibility, utilizing both glycolysis and oxidative phosphorylation, thus providing a relevant host for investigating cancer cell metabolism and lipid signaling in a lung cancer background.
AGPAT2 catalyzes the acylation of LPA to PA, a precursor for all glycerolipids, including diacylglycerol (DAG), triacylglycerol, and major phospholipid classes such as phosphatidylcholine and phosphatidylethanolamine. Its expression is transcriptionally regulated by PPARG and SREBF1, and it operates upstream of LPIN1 (converts PA to DAG) and DGAT1 (converts DAG to triacylglycerol). The PA generated by AGPAT2 also serves as a lipid second messenger, activating mTORC1 and Raf kinase pathways. Additionally, AGPAT2 functionally interacts with AGPAT1 and GPAT enzymes, orchestrating glycerolipid synthesis and lipid droplet biogenesis.
In A-549 cells, AGPAT2 knockout disrupts de novo glycerolipid synthesis, leading to reduced membrane phospholipid and triacylglycerol pools. This deficiency impairs membrane biogenesis, lipid droplet formation, and mTORC1-mediated proliferative signaling, which is frequently dysregulated in lung adenocarcinoma. The model also recapitulates aspects of congenital generalized lipodystrophy type 1, where AGPAT2 mutations cause severe lipoatrophy, insulin resistance, and metabolic syndrome, enabling cross-tissue investigations of AGPAT2 function beyond adipocytes.
These polyclonal knockout cells are ideally suited for diverse applications, including lipidomics by mass spectrometry, quantitative triglyceride assays, Oil Red O staining for lipid droplet visualization, and Seahorse metabolic flux analysis to monitor energy metabolism. Western blotting for phospho-p70 S6K assesses mTORC1 signaling, while RT-qPCR confirms AGPAT2 transcript ablation. Cell proliferation assays can reveal growth defects caused by lipid scarcity. The model supports drug target validation, adipogenesis research, and cancer metabolism studies. For further information, please contact Ascent Research.
