The BDH2 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HAP1 cells with targeted disruption of the BDH2 gene. This pooled knockout format provides a genetically heterogeneous loss-of-function model, enabling robust analysis of BDH2-dependent phenotypes without clonal expansion bias.
The HAP1 cell line is a near-haploid human cell model derived from the KBM-7 chronic myeloid leukemia cell line. Its near-haploid karyotype and fibroblast-like morphology make it an ideal platform for knockout, genetic interaction, and functional genomic studies. HAP1 cells are widely employed in arrayed and pooled CRISPR screens, as the single allelic copy simplifies loss-of-function analysis and reduces genetic redundancy.
BDH2 encodes a mitochondrial dehydrogenase that catalyzes the NAD+-dependent oxidation of 3-hydroxybutyrate to acetoacetate, a key step in ketone body utilization and mitochondrial energy metabolism. In addition, BDH2 synthesizes the siderophore 2,5-dihydroxybenzoic acid (2,5-DHBA), which chelates ferric iron to regulate intracellular iron homeostasis and mitigate oxidative stress. BDH2 expression is upregulated by PPAR?? signaling during fasting and is suppressed by HIF-1?? under hypoxia. Downstream, BDH2 activity influences the mitochondrial NAD+/NADH ratio, acetoacetate production, and iron availability through 2,5-DHBA-mediated chelation. BDH2 functionally interacts with mitochondrial electron transport chain components and iron regulatory proteins such as ferritin, positioning it at the intersection of energy metabolism and iron regulation.
In the HAP1 background, BDH2 knockout provides a controlled system to dissect mitochondrial ketone body oxidation and iron-siderophore biology. The near-haploid genome minimizes confounding gene copies, making this model especially suited for studying metabolic reprogramming in cancer cells and for high-throughput screening of chemical or genetic modulators. Disruption of BDH2 in HAP1 cells allows direct assessment of altered ketone body flux, iron accumulation, and redox balance in a genetically simplified context.
This BDH2 knockout cell pool is applicable to a range of experimental workflows. Researchers can employ Western blotting and RT-qPCR to confirm BDH2 loss, quantify ??-hydroxybutyrate and acetoacetate levels to evaluate ketone metabolism, and use fluorescent probes such as FerroOrange to detect intracellular iron changes. Mitochondrial function can be assessed via Seahorse respirometry, while oxidative stress can be monitored with DCFDA/H2DCFDA-based ROS assays. The model supports investigations into iron overload disorders, neurodegeneration with brain iron accumulation, and cancer metabolic adaptation. For further information, please contact Ascent Research.
