The GPD1L Knockout HT29 Polyclonal Cells are a polyclonal population of HT29 human colorectal adenocarcinoma cells in which the GPD1L gene has been disrupted using CRISPR/Cas9 gene editing. This targeted disruption generates a loss-of-function model, eliminating functional GPD1L expression across a genetically diverse cell pool. The polyclonal format avoids clonal artifacts and provides a robust system for studying GPD1L-dependent phenotypes in a context that mirrors tumor heterogeneity.
HT29 is a well-established human colorectal adenocarcinoma cell line, originally derived from a primary tumor. These adherent epithelial cells bear oncogenic mutations driving constitutive proliferation and survival signaling, and they exhibit a strong reliance on glycolytic metabolism even in normoxia. HT29 cells adapt to hypoxic microenvironments, making them an excellent model for investigating metabolic reprogramming and hypoxia-driven signaling in colorectal cancer.
GPD1L encodes glycerol-3-phosphate dehydrogenase 1-like, an enzyme that catalyzes the conversion of dihydroxyacetone phosphate (DHAP) to glycerol-3-phosphate, coupled with the oxidation of NADH to NAD+. This reaction is essential for the glycerol phosphate shuttle, which transfers cytosolic reducing equivalents into mitochondria and regenerates NAD+ to sustain glycolytic flux. GPD1L is transcriptionally regulated by HIF-1?? under hypoxia and physically interacts with the sodium channel SCN5A. Downstream, GPD1L modulates the NAD+/NADH ratio, influencing SIRT1 activity, ROS production, and AMPK signaling, thereby integrating metabolic state with energy-sensing and redox pathways.
In HT29 colorectal adenocarcinoma cells, GPD1L is thought to support the Warburg effect by maintaining glycolysis and redox balance under hypoxia. Disruption of GPD1L in this polyclonal knockout model is expected to impair glycerol phosphate shuttle activity, leading to a reduced NAD+/NADH ratio, diminished glycolytic capacity, and increased oxidative stress. These metabolic alterations render cells more susceptible to glycolysis inhibitors and nutrient deprivation, offering a powerful tool to study metabolic dependencies and therapeutic vulnerabilities in colorectal cancer.
This knockout product is suited for metabolic profiling with Seahorse glycolysis stress tests, NAD+/NADH quantification, and Western blotting for HIF-1?? and glycolytic enzymes. It also enables flow cytometry-based apoptosis assays under glucose deprivation, drug sensitivity screens with glycolysis inhibitors, and studies of GPD1L?CSCN5A interactions. The polyclonal format facilitates pooled phenotypic analyses relevant to tumor biology. For further information, please contact Ascent Research.
