The GNPAT Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, featuring targeted disruption of the GNPAT gene. This polyclonal pool encompasses a heterogeneous mix of edited cells, enabling functional study of GNPAT loss in a relevant epithelial background without clonal selection constraints. The knockout is achieved through CRISPR/Cas9-mediated gene disruption, abolishing expression of glyceronephosphate O-acyltransferase, the peroxisomal enzyme that initiates ether lipid biosynthesis. This polyclonal model is optimized for investigating peroxisomal lipid metabolism and its consequences in intestinal epithelial cells.
The HT29 parental line is an established model of colorectal adenocarcinoma with epithelial morphology, extensively employed in intestinal biology, drug transport, and metabolism research. HT29 cells maintain key intestinal epithelial characteristics and can undergo enterocytic differentiation, offering a physiologically relevant system. Their robust growth and well-documented genetic profile ensure experimental reproducibility. The line is particularly valuable for studying drug metabolism and pharmacokinetic properties, owing to its expression of metabolic enzymes and transporters. These features make HT29 an ideal host for dissecting GNPAT-dependent metabolic pathways.
GNPAT encodes glyceronephosphate O-acyltransferase, which catalyzes the first step in ether lipid biosynthesis within peroxisomes: the acylation of dihydroxyacetone phosphate (DHAP) with a long-chain acyl-CoA. This reaction is upstream of alkylglycerone phosphate synthase (AGPS), with which GNPAT functionally interacts to ensure efficient plasmalogen production. The pathway is regulated by PPAR-alpha and fatty acid levels, linking peroxisomal function to systemic lipid status. Downstream products include plasmalogens such as phosphatidylethanolamine plasmalogen and other ether-linked phospholipids, which are essential for membrane architecture and signaling. Loss of GNPAT disrupts this cascade, impairing plasmalogen synthesis and altering membrane properties.
In the colorectal adenocarcinoma context, GNPAT knockout provides a platform to elucidate the roles of plasmalogens in intestinal epithelial homeostasis and tumor cell metabolism. Altered peroxisomal lipid metabolism is implicated in cancer progression, and this model enables dissection of how ether lipid deficiency impacts proliferation, differentiation, and drug sensitivity. It also serves as a cellular system for modeling rhizomelic chondrodysplasia punctata type 2, a severe peroxisomal disorder caused by GNPAT mutations. Combining HT29??s intestinal epithelial features with GNPAT disruption yields a powerful tool for translational research.
Typical research applications include detailed investigation of ether lipid metabolism, peroxisomal biogenesis, and membrane biology through assays such as mass spectrometry-based lipid profiling, western blotting for GNPAT protein, RT-qPCR for transcript analysis, and immunofluorescence for peroxisomal localization. This polyclonal knockout product is also suitable for metabolic flux analysis, cell viability studies, and drug sensitivity screens to assess therapeutic responses under plasmalogen-deficient conditions. For further technical details or to request a quotation, please contact Ascent Research.
