AGPAT4 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted AGPAT4 disruption in HeLa cells. This loss-of-function model enables study of AGPAT4-dependent processes in human cervical adenocarcinoma. The polyclonal format provides a heterogeneous cell pool for population-level analyses, avoiding clonal artifacts. These cells facilitate dissection of AGPAT4??s role in glycerolipid metabolism and downstream cellular phenotypes.
HeLa cells are an immortalized cervical adenocarcinoma line, HPV18-positive and p53-deficient. Widely used in cancer research, they exhibit epithelial morphology and stable in culture, with a well-characterized transcriptome. The AGPAT4 knockout in this background offers a tool to study lipid metabolic adaptations in a cancer-relevant epithelial system.
AGPAT4 catalyzes the acylation of lysophosphatidic acid (LPA) to phosphatidic acid (PA) in the endoplasmic reticulum, a critical step in de novo glycerolipid biosynthesis. This reaction controls fatty acid flux into triacylglycerols and membrane phospholipids, influencing lipid droplet formation and PA signaling. AGPAT4 is transcriptionally regulated by SREBP1 and ChREBP downstream of insulin and PPAR??, and functionally interacts with Lipin phosphatidate phosphatases and acyl-CoA synthetases. Its activity promotes PA accumulation, fueling triglyceride synthesis via Lipin?Cdiacylglycerol?CDGAT and phospholipid remodeling. Disruption thus alters lipid storage and signaling, enabling dissection of these interconnected networks.
In HeLa cells, which exhibit elevated lipogenesis and lipid droplet accumulation, AGPAT4 knockout is relevant for probing cancer lipid metabolism. By ablating AGPAT4, researchers can examine how altered PA levels affect lipid droplet dynamics, membrane phospholipid composition, and cancer cell growth. This model may help clarify AGPAT4??s role in tumor lipid metabolism, an area linked to intellectual disability and cancer. Metabolic flux analyses in this p53-deficient, HPV-positive background can map carbon allocation toward neutral lipids versus phospholipids.
These polyclonal cells enable a range of assays: lipidomic profiling by LC-MS/MS to quantify glycerolipid changes; Oil Red O or BODIPY staining for neutral lipids; Western blotting for lipogenic enzymes; [^3H]-palmitate incorporation to measure triglyceride synthesis; Seahorse flux analysis for metabolic shifts; and proliferation assays for cancer cell fitness. For additional details or custom assay development, contact Ascent Research.
