The AGPAT1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited HT29 human colon adenocarcinoma cell population with disrupted AGPAT1 gene function. This polyclonal knockout model enables loss-of-function studies of 1-acylglycerol-3-phosphate O-acyltransferase 1 (AGPAT1), a key enzyme in de novo phospholipid and triglyceride synthesis, by abolishing the conversion of lysophosphatidic acid (LPA) to phosphatidic acid (PA). The product thus provides a valuable tool for investigating lipid metabolic reprogramming in epithelial cancer biology.
The HT29 host cell line, derived from a human colorectal adenocarcinoma, retains intestinal epithelial characteristics and can undergo differentiation into enterocytic lineages under controlled conditions. Widely used in intestinal barrier function, nutrient transport, and oncogenic signaling studies, HT29 cells form polarized monolayers and serve as a clinically relevant model for colorectal cancer research. This background makes them particularly suitable for exploring how lipid metabolism perturbations influence epithelial polarity, membrane dynamics, and tumor-associated processes.
AGPAT1 catalyzes the acylation of LPA to PA, a central reaction in the Kennedy pathway. PA functions as a metabolic hub for phospholipid and triglyceride production and directly activates mTORC1, coupling nutrient status to anabolic growth. AGPAT1 expression is transcriptionally controlled by SREBP1 and PPAR??, integrating insulin signaling. The enzyme interacts with lipin-1 and LPIN2, which convert PA to diacylglycerol, and cooperates with GPAT in upstream acylation steps. Disruption of AGPAT1 therefore depletes PA levels, attenuates mTOR signaling, and alters glycerolipid homeostasis.
In HT29 colon cancer cells, AGPAT1 knockout enables dissection of phospholipid remodeling effects on malignant behavior. Altered PA pools may disrupt membrane biophysics, vesicular trafficking, and mTORC1-driven proliferation. Given the frequent rewiring of lipid metabolism in colorectal tumors, AGPAT1 loss shifts cellular lipid profiles and could reveal metabolic sensitivities. This model is particularly valuable for studying the interplay between lipid signaling and oncogenic pathways such as Wnt/??-catenin.
Recommended applications include western blotting and RT-qPCR for AGPAT1 expression validation, lipidomic profiling to map lipid alterations, and metabolic assays to quantify triglyceride synthesis. Functional readouts encompass proliferation, migration, and invasion assays, while phospho-signaling analyses track mTORC1 pathway activity. Barrier integrity experiments in polarized HT29 monolayers further assess AGPAT1??s role in epithelial homeostasis. These capabilities support drug screening for metabolic enzyme inhibitors and mechanistic studies in cancer lipid metabolism. For additional technical information, please contact Ascent Research.
