The BDH1 Knockout HT29 Polyclonal Cells are a heterogeneous population of HT29 colorectal adenocarcinoma cells with CRISPR/Cas9-mediated disruption of the BDH1 gene. This polyclonal pool enables functional studies of BDH1 in colon cancer biology without clonal selection, retaining editing-associated genetic diversity that better reflects tumor heterogeneity. Supplied as early-passage, ready-to-use knockout cells, they facilitate investigation of ketone body utilization in cancer metabolism.
The HT29 host cell line is a well-established human colorectal adenocarcinoma epithelial model, carrying oncogenic mutations in APC, TP53, KRAS, and BRAF, with microsatellite stability. Widely used in cancer biology and drug discovery, HT29 cells form adherent monolayers and represent a clinically relevant system for studying signaling and metabolic adaptations. BDH1 knockout in this background allows assessment of ketone body metabolic flux in a genetically defined cancer context.
BDH1 (D-beta-hydroxybutyrate dehydrogenase 1) is a mitochondrial enzyme catalyzing NAD+-dependent oxidation of (R)-3-hydroxybutyrate to acetoacetate, the first step in ketone body utilization. Its activity is regulated by PPARA, HNF4A, and glucocorticoid receptor, and is induced by fasting or ketogenic stimuli. BDH1 interacts with mitochondrial trifunctional protein and partners with HMGCS2, OXCT1, and ACAT1 in ketolysis. The acetoacetate product is further metabolized to acetyl-CoA, driving the TCA cycle and ATP production, linking ketone catabolism to mitochondrial respiration in nutrient-depleted conditions.
In HT29 cells, BDH1 disruption provides a model to study ketone body metabolism in tumor survival and proliferation. Colorectal tumors often face nutrient fluctuations, and ketone body utilization may confer a metabolic advantage during fasting or calorie restriction. BDH1 knockout enables investigation of how loss of this ketolytic enzyme affects bioenergetics, redox balance, and sensitivity to metabolic inhibitors, making it relevant for exploring metabolic reprogramming and therapeutic targeting in colorectal carcinoma and other ketone-utilizing cancers.
Applications include metabolic flux analysis with isotope-labeled substrates, Seahorse respirometry, and viability assays under glucose-depleted or ketone-rich conditions. Western blotting and RT-qPCR confirm BDH1 disruption and assess compensation by BDH2 or OXCT1. Additional assays such as ketone body quantification and immunofluorescence support phenotypic characterization. This knockout model is suitable for studies of metabolic acidosis, ketone body utilization disorders, and cancer metabolism. For detailed technical specifications or to request a quote, please contact Ascent Research.
