The GPD2 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population targeting the GPD2 gene within the HT29 colorectal adenocarcinoma background. This heterogeneous pool contains numerous loss-of-function disruptions, avoiding the need for single-cell cloning and enabling the study of GPD2 deficiency in a tumorigenic epithelial context. The polyclonal format captures a range of knockout phenotypes, making it suitable for direct functional assays including metabolic flux analysis, signaling studies, and drug response profiling.
The HT29 cell line is a well-established model of human colorectal adenocarcinoma, derived from a 44-year-old female patient. Characterized by TP53 and APC mutations and microsatellite stability, HT29 cells recapitulate key oncogenic features of non-hypermutated colorectal tumors. These adherent epithelial cells maintain enterocyte differentiation potential, providing a relevant system for investigating metabolic gene disruptions in intestinal cancer. This genetic landscape is essential for examining how GPD2 loss alters cancer cell metabolism within a defined oncogenic context.
GPD2 encodes mitochondrial glycerol-3-phosphate dehydrogenase (mGPDH), which catalyzes the oxidation of glycerol-3-phosphate to DHAP in the glycerol phosphate shuttle, coupling cytosolic glycolysis to oxidative phosphorylation. This reaction transfers reducing equivalents to ubiquinone, generating FADH? and maintaining NAD?/NADH balance. GPD2 is regulated by insulin, glucose, PPAR??, and PGC-1??, and its activity modulates ATP synthesis, ROS production, and DHAP levels. The enzyme interacts with ubiquinone and mitochondrial respiratory chain components, integrating lipid and carbohydrate metabolism at the inner mitochondrial membrane.
In HT29 colorectal adenocarcinoma cells, GPD2 knockout disrupts the glycerol phosphate shuttle, impairing the transfer of reducing equivalents from cytosolic NADH to the electron transport chain. This perturbation leads to altered redox homeostasis, reduced mitochondrial ATP production, and compensatory shifts in glycolysis and lipid metabolism??key aspects of metabolic reprogramming in cancer. The TP53/APC mutant background of HT29 cells provides an ideal platform to investigate how oncogenic signaling converges with mitochondrial shuttle dependencies, potentially revealing metabolic vulnerabilities in colorectal tumors.
This knockout model supports investigations into mitochondrial metabolism and cancer biology through a variety of assays, including GPD2 activity measurements, Seahorse respirometry, lactate and ATP quantification, ROS detection, western blotting, and RT-qPCR. It enables studies of redox balance, glycolytic flux, and metabolic adaptation, as well as drug screening for inhibitory compounds targeting mitochondrial shuttles. The cells are also valuable for modeling type 2 diabetes and obesity-related metabolic dysfunction in a cancer-relevant context. For further details, contact Ascent Research.
