The EHHADH Knockout 786-O Polyclonal Cells product provides a polyclonal population of human 786-O renal cell carcinoma cells that have undergone CRISPR/Cas9-mediated disruption of the EHHADH gene. This polyclonal knockout format preserves the intrinsic genetic heterogeneity of the parental line while effectively ablating target gene function across the bulk culture, serving as a versatile tool for loss-of-function studies without clonal selection biases.
The parental 786-O cell line is a widely used model of clear cell renal cell carcinoma (ccRCC) derived from a primary renal adenocarcinoma. These adherent epithelial cells harbor a biallelic VHL tumor suppressor mutation, resulting in constitutive HIF stabilization and a metabolic shift toward aerobic glycolysis and lipid accumulation. This genetic background makes 786-O cells particularly suited for studying lipid metabolism and oncogenic signaling.
EHHADH encodes the peroxisomal L-bifunctional protein, which catalyzes the enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase steps of straight-chain fatty acid ??-oxidation. Its expression is upregulated by the lipid-sensing nuclear receptors PPARA and PPARG. After PEX5-mediated peroxisomal import, EHHADH functions downstream of ACOX1 and collaborates with HSD17B4 and other components such as SCP2 and ACAA1 to shorten very long-chain fatty acids, producing acetyl-CoA, NADH, and acyl-CoA intermediates. This pathway is essential for cellular energy homeostasis and lipid metabolism.
In VHL-mutant 786-O cells, loss of EHHADH disrupts peroxisomal ??-oxidation, leading to accumulation of very long-chain fatty acids and a forced reliance on alternative metabolic pathways. This model recapitulates metabolic features of peroxisomal disorders within a renal cancer context, enabling investigation of how peroxisomal dysfunction interacts with oncogenic signaling and HIF-driven metabolic reprogramming. It is a powerful system to dissect PPAR??-mediated transcriptional networks and identify metabolic vulnerabilities.
This polyclonal knockout cell pool is suitable for a broad range of experimental readouts. Western blotting and RT-qPCR validate EHHADH knockout and monitor PPAR??-responsive genes. Fatty acid oxidation assays and metabolomic profiling quantify metabolic flux and very long-chain fatty acid accumulation. Oil Red O staining and Seahorse analysis assess lipid storage and cellular respiration. Immunofluorescence localizes peroxisomal markers. These cells also support high-throughput screening for peroxisomal metabolism modulators and functional studies of peroxisomal disorders. For further information, please contact Ascent Research.
