The ECHDC3 Knockout 786-O Polyclonal Cells product offers a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 786-O renal clear cell carcinoma line, featuring targeted disruption of the ECHDC3 gene. This loss-of-function model enables systematic investigation of enoyl-CoA hydratase domain-containing protein 3 in the context of cancer metabolism.
The 786-O cell line is a widely used model of human clear cell renal cell carcinoma (ccRCC). It harbors a truncating mutation in the VHL tumor suppressor gene, leading to constitutive stabilization of hypoxia-inducible factors HIF-1?? and HIF-2?? under normoxic conditions. This pseudohypoxic state drives transcriptional programs that promote angiogenesis, glycolytic metabolism, and lipid accumulation, closely reflecting the metabolic adaptations observed in ccRCC tumors.
ECHDC3 encodes a mitochondrial enzyme that catalyzes the hydration of trans-2-enoyl-CoA intermediates to L-3-hydroxyacyl-CoA during the second step of the fatty acid ??-oxidation spiral. The protein operates within a multienzyme complex that includes very long-chain acyl-CoA dehydrogenase (ACADVL) and the trifunctional protein subunits HADHA and HADHB, acting downstream of the carnitine palmitoyltransferases CPT1 and CPT2. This reaction generates NADH and FADH?, which transfer electrons to the respiratory chain, and yields acetyl-CoA for the tricarboxylic acid cycle, thereby contributing to ATP production. ECHDC3 expression is regulated by the nuclear receptors PPAR?? and the coactivator PGC-1??, key orchestrators of mitochondrial biogenesis and fatty acid oxidation, and is additionally modulated by AMPK- and insulin-dependent signaling pathways, linking nutrient availability to lipid catabolic activity.
In VHL-deficient 786-O cells, fatty acid oxidation may provide an important source of energy to support proliferation under metabolic stress. Disruption of ECHDC3 impairs this catabolic pathway, potentially limiting ATP generation from lipids and forcing metabolic rewiring. This knockout model thus allows researchers to probe the role of mitochondrial ??-oxidation in ccRCC and to identify metabolic vulnerabilities that could be exploited therapeutically.
Typical experimental workflows include validation of knockout efficiency by Western blot and RT-qPCR, functional assessment of fatty acid oxidation using radiolabeled palmitate assays, and metabolic flux analysis with Seahorse XF analyzers (OCR/ECAR). Complementary approaches such as ATP luminescence assays, lipidomic profiling by LC-MS, and cell proliferation or migration assays can delineate the broader consequences of ECHDC3 loss. Moreover, examination of HIF target gene expression (e.g., VEGF, GLUT1) may uncover interactions between hypoxia signaling and lipid metabolism. This product is ideally suited for investigating metabolic reprogramming in renal cell carcinoma, evaluating therapeutic strategies targeting energy metabolism, and dissecting the regulatory networks linking lipid catabolism to oncogenic signaling. For further technical information, please contact Ascent Research.
