The HADHB Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the HADHB gene. This product provides a heterogeneous pool of HAP1 cells carrying diverse loss-of-function alleles, ensuring robust gene disruption without clonal isolation. The polyclonal format maintains population-level biological complexity and is well-suited for metabolic studies requiring near-complete elimination of HADHB function.
The host HAP1 line is a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia cells. Its haploid genome simplifies gene knockout by requiring disruption of only a single allele, eliminating the potential for heterozygous compensation. HAP1 cells retain key metabolic pathways and are widely used in genetic screens, drug discovery, and functional genomics, providing a clean experimental platform for investigating mitochondrial fatty acid oxidation.
HADHB encodes the beta subunit of the mitochondrial trifunctional protein (MTP), a multienzyme complex that catalyzes the hydration of long-chain enoyl-CoAs to 3-hydroxyacyl-CoAs and their reduction to 3-oxoacyl-CoAs, essential steps in long-chain fatty acid beta-oxidation. HADHB interacts physically with the alpha subunit HADHA to form the active MTP complex. Its expression is regulated by metabolic sensors including PPARalpha, PGC-1alpha, AMPK, and insulin signaling. The reaction products acetyl-CoA, NADH, and FADH2 fuel the TCA cycle and electron transport chain, while acylcarnitines are generated as transport intermediates. HADHB also functionally cooperates with ECHS1 and ACADVL within the beta-oxidation pathway. Disruption of HADHB impairs fatty acid oxidation and disrupts mitochondrial energy homeostasis.
In the HAP1 near-haploid context, HADHB knockout creates a definitive loss-of-function model for mitochondrial trifunctional protein deficiency. The single-allele disruption ensures that the polyclonal population faithfully recapitulates metabolic defects observed in human diseases such as long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency. Impaired fatty acid utilization leads to energy depletion and lipid accumulation, which underlie myopathy, neuropathy, and sudden infant death syndrome. This model enables detailed investigation of disease mechanisms and metabolic vulnerabilities.
This knockout cell population supports diverse applications including metabolic disease modeling, drug screening for modulators of fatty acid oxidation, and functional genomics. Representative assays include fatty acid oxidation measurements, metabolic flux analysis using labeled substrates, lipidomic profiling of acylcarnitines, Western blotting for HADHB confirmation, immunofluorescence to examine mitochondrial morphology, and viability assays under metabolic stress such as glucose deprivation or oxidative challenge. The polyclonal cells are also suitable for CRISPR-based genetic interaction studies. For additional information or custom requests, please contact Ascent Research.
