The AGPAT3 Knockout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of HT29 cells with targeted disruption of the AGPAT3 gene, providing a loss-of-function model for studying the roles of AGPAT3 in lipid metabolism and signaling. This polyclonal knockout pool is generated using CRISPR/Cas9-mediated gene disruption and is ideal for assays that require a heterogeneous population, such as pooled screening or studies of heterogeneous responses.
The HT29 host cell line originates from a primary colorectal adenocarcinoma of a female patient and exhibits epithelial morphology with the capacity for enterocytic differentiation and mucin secretion. These cells serve as a well-established in vitro model for colorectal cancer research, particularly for studying intestinal epithelial biology, mucin production, and tumor cell differentiation. HT29 cells are widely used for investigating signal transduction, drug responses, and metabolic adaptations in colon cancer.
AGPAT3 encodes a 1-acylglycerol-3-phosphate O-acyltransferase that catalyzes the conversion of lysophosphatidic acid (LPA) to phosphatidic acid (PA), a central intermediate in glycerolipid and glycerophospholipid biosynthesis. AGPAT3 activity is transcriptionally regulated by adipogenic transcription factors such as PPAR??, SREBP-1c, and C/EBP??, and responds to insulin signaling. The produced PA serves as a precursor for diacylglycerol, triglycerides, and phospholipids, and also functions as a lipid second messenger that activates mTOR and PKC signaling pathways. AGPAT3 interacts with Lipin1 and ER membrane proteins, and is implicated in lipid droplet formation.
Disruption of AGPAT3 in the HT29 colorectal cancer background enables investigation of phosphatidic acid-dependent signaling and lipid metabolic reprogramming in intestinal tumor cells. Because PA is a potent activator of mTOR and PKC, AGPAT3 loss likely attenuates these pro-proliferative and survival cues, potentially impacting tumor growth, differentiation, and migration. Moreover, HT29 cells can undergo enterocytic differentiation and produce mucins, and AGPAT3-mediated lipid metabolism may regulate these processes. This knockout model thus provides a relevant system to dissect how altered glycerolipid biosynthesis influences colorectal cancer cell behavior and metabolic vulnerability.
Researchers can employ this polyclonal knockout pool to study the impact of AGPAT3 loss on lipid metabolism, phosphatidic acid signaling, and cellular phenotypes. Suitable assays include lipid profiling by LC-MS, triglyceride quantification, Oil Red O staining for lipid droplet assessment, Western blotting and RT-qPCR for verification of knockout and downstream targets, cell proliferation (MTT) and migration/invasion assays, and phospho-kinase arrays to probe mTOR/PKC pathway activity. Transcriptome-wide analysis via RNA-seq can reveal AGPAT3-dependent gene expression changes. This product is well-suited for drug target screening in metabolic and oncological contexts. For further details, please contact Ascent Research.
