The AGPAT5 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population in which the AGPAT5 gene has been disrupted. This loss-of-function model is derived from the HT-29 colorectal adenocarcinoma line and enables the systematic study of AGPAT5-mediated lipid metabolism and signaling.
The HT-29 cell line was established from a primary colorectal adenocarcinoma and is extensively used as an intestinal epithelial model. These cells are capable of enterocytic differentiation under appropriate conditions and serve as a standard system for investigating drug absorption, epithelial biology, and colorectal cancer mechanisms. Their well-documented growth characteristics provide a reliable host for gene-editing studies.
AGPAT5 (1-acylglycerol-3-phosphate O-acyltransferase 5) catalyzes the acylation of lysophosphatidic acid (LPA) to produce phosphatidic acid (PA), a key intermediate in glycerophospholipid and triacylglycerol synthesis. The enzyme is transcriptionally activated by SREBP1c and PPARgamma and is responsive to insulin, integrating hormonal and nutritional cues. Using LPA and acyl-CoA as substrates, AGPAT5 generates PA, which not only serves as a precursor for diacylglycerol (DAG) but also functions as a lipid second messenger. PA directly activates mTORC1, linking lipid metabolism to cell growth and proliferation. Disruption of AGPAT5 therefore impairs both membrane lipid biogenesis and mTOR-mediated signaling cascades.
In the HT-29 colorectal adenocarcinoma context, knockout of AGPAT5 disrupts de novo phospholipid synthesis, altering membrane composition and attenuating lipid-derived signals. This can compromise the proliferation and survival of tumor cells that heavily rely on lipid biosynthesis. The model enables detailed investigation of how colorectal cancer cells adapt to metabolic stress and the role of lipid signaling in tumor maintenance. It also offers a platform to explore the intersection between glycerolipid metabolism and oncogenic pathways.
This polyclonal knockout population is suitable for lipidomic analyses, cell proliferation assays, and metabolic tracing with [14C]-glycerol. Signaling changes can be assessed via western blot for phospho-S6K1, a readout of mTORC1 activity, while lipid accumulation is visualized by Oil Red O staining. RT-qPCR of lipogenic genes can further characterize transcriptional responses. These applications support research into tumor metabolism, drug response, and intestinal nutrient handling. For more details, please contact Ascent Research.
