The HADH Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that disrupts expression of the HADH gene in the near-haploid HAP1 human cell line. This product provides a heterogeneous pool of cells with targeted HADH disruption, enabling bulk analysis of loss-of-function phenotypes without requiring clonal selection. HADH encodes mitochondrial hydroxyacyl-CoA dehydrogenase, a critical enzyme in short- and medium-chain fatty acid ??-oxidation. By ablating HADH, researchers can investigate its role in mitochondrial metabolism, energy homeostasis, and disease-associated pathways.
HAP1 cells are derived from KBM-7 chronic myeloid leukemia blast crisis cells and exhibit a stable near-haploid karyotype. This genetic simplicity removes confounding effects of diploidy, ensuring clear genotype-to-phenotype linkages. HAP1??s rapid proliferation, ease of culture, and myeloid leukemic origin make it a powerful platform for CRISPR-based functional genomics studies in cancer metabolism, hematological disorders, and metabolic signaling.
HADH catalyzes the third step of mitochondrial ??-oxidation, oxidizing medium- and short-chain 3-hydroxyacyl-CoAs and generating NADH. It lies downstream of ACADM/ACADS and upstream of ACAT1. Transcriptional control involves PPARA, PGC-1??, HNF4A, and CREB1. HADH interacts with ETFA/ETFB, GLUD1, and MRPL12, functioning as a homodimer. Through NADH production, it regulates the ATP/ADP ratio and, in ??-cells, insulin secretion.
Knockout of HADH in HAP1 cells leads to defective fatty acid oxidation, accumulating medium- and short-chain acylcarnitines and lowering ATP levels. This mimics metabolic derangements seen in congenital hyperinsulinemic hypoglycemia and fatty acid oxidation disorders, providing a relevant in vitro model. The polyclonal knockout in a near-haploid background yields a reproducible, penetrant phenotype ideal for dissecting HADH-dependent metabolic reprogramming.
These cells are valuable for functional genomics, metabolic disease modeling, and drug screening. Researchers can perform acylcarnitine profiling, 14C-palmitate oxidation assays, Seahorse mitochondrial stress tests, and intracellular ATP measurements to characterize HADH-dependent pathways and screen for pharmacological modulators. For further technical details, please contact Ascent Research.
