ECHDC2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human cell line. These cells contain a heterogeneous mixture of ECHDC2 gene disruptions, created without clonal isolation, thereby preserving genetic diversity and reducing clone-specific artifacts. The polyclonal format ensures that functional studies average out the effects of any single-gene edit, providing a robust loss-of-function model. The cells are supplied as a live population and are intended for research use only.
HAP1 is a near-haploid cell line derived from the male chronic myeloid leukemia line KBM-7, exhibiting an adherent fibroblast-like morphology. Its haploid karyotype simplifies genetic manipulation, requiring disruption of a single allele to achieve functional knockout. HAP1 cells are extensively used in genetic screens and functional genomics due to their stable growth and facile engineering. They retain active mitochondrial fatty acid oxidation pathways, making them suitable for studying metabolic gene function.
ECHDC2 encodes a mitochondrial enoyl-CoA hydratase predicted to participate in the beta-oxidation of unsaturated fatty acids. Expression of ECHDC2 is regulated by the nuclear receptors PPARA and PPARG, which are master regulators of lipid catabolism. Inside mitochondria, ECHDC2 hydrates enoyl-CoA intermediates downstream of ACADVL, in concert with the trifunctional protein subunits HADHA and HADHB, and upstream of ECHS1 and ACAA2. Disruption of ECHDC2 interrupts this pathway, impairing the conversion of unsaturated fatty acids into acetyl-CoA and ketone bodies and potentially leading to acyl-CoA accumulation and metabolic stress.
Given the leukemic origin of HAP1 cells, ECHDC2 knockout in this background enables exploration of lipid metabolism rewiring in cancer. ECHDC2 has been linked to colorectal cancer susceptibility in GWAS, suggesting that its loss may modulate metabolic phenotypes relevant to oncogenesis. The HAP1 platform permits integration of this genetic lesion with high-throughput screening to assess its effect on cell fitness, drug sensitivity, and metabolic flux.
These polyclonal knockout cells are suitable for a range of metabolic studies, including Seahorse-based respirometry to measure mitochondrial function, radiolabeled fatty acid oxidation assays, and western blotting for pathway analysis. They can be employed to validate targets from genetic screens, dissect PPAR signaling, and investigate the role of ECHDC2 in cancer metabolism. Researchers may also utilize the cells for immunofluorescence microscopy or mitochondrial isolation to assess enzyme localization and activity. For further information, please contact Ascent Research.
