The ECI1 Knockout 786-O Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population derived from the human renal clear cell carcinoma line 786-O, featuring disruption of the ECI1 gene encoding enoyl-CoA delta isomerase 1. This product provides a heterogeneous cell pool with diverse loss-of-function mutations at the ECI1 locus, enabling functional studies of unsaturated fatty acid oxidation in a VHL-null ccRCC background without single-cell cloning, thus maintaining biological variability while achieving robust gene disruption.
The parental 786-O cell line is a widely used model of clear cell renal cell carcinoma (ccRCC) that harbors biallelic VHL inactivation, resulting in constitutive stabilization of HIF-1?? and pseudohypoxic signaling. This VHL-deficient background reproduces key metabolic reprogramming features of ccRCC, including heightened glycolysis, glutamine-dependent reductive carboxylation, and altered lipid metabolism, making it ideal for investigating how ECI1 deficiency interacts with HIF-driven metabolic rewiring.
ECI1 catalyzes the isomerization of cis-3-enoyl-CoA to trans-2-enoyl-CoA esters, a critical step in mitochondrial ??-oxidation of unsaturated fatty acids. This reaction is required for the trifunctional protein complex (comprising HADHA and HADHB) to complete fatty acid degradation. ECI1 is transcriptionally regulated by PPAR?? and PGC-1??, and its expression is modulated by HIF-1?? under hypoxia. The enzyme interacts with ACADVL and other ??-oxidation components to generate acetyl-CoA, TCA cycle intermediates, and ATP. Disruption of ECI1 blocks efficient unsaturated fatty acid utilization, potentially causing lipid intermediate accumulation and metabolic stress.
In the VHL-null 786-O ccRCC model, ECI1 loss likely exacerbates the inherent mitochondrial fatty acid oxidation deficiency observed in clear cell tumors. ccRCC cells rely heavily on fatty acid uptake and lipid droplet storage, yet often display reduced ??-oxidation capacity. ECI1 knockout may further impair energy production from unsaturated fatty acids, forcing a metabolic shift toward glycolysis and glutaminolysis. This vulnerability could be targeted therapeutically, positioning these polyclonal knockout cells as a tool for identifying synthetic lethal interactions in renal cancer metabolism.
These polyclonal knockout cells are suited for metabolic and oncological studies, including fatty acid oxidation flux assays with 14C-palmitate, mitochondrial respiration measurements (Seahorse), and metabolomics profiling of lipid intermediates and TCA cycle metabolites. RT-qPCR and western blotting can confirm ECI1 disruption and compensatory enzyme expression. Additional applications encompass cell proliferation assays under lipid-rich conditions, drug sensitivity screens, and investigation of PPAR??/PGC-1??/HIF-1?? crosstalk. For further information or custom knockout cell generation services, please contact Ascent Research.
