The ECHS1 Knockout 786-O Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in the 786-O human clear cell renal cell carcinoma (ccRCC) line, featuring targeted disruption of the ECHS1 gene. This gene encodes mitochondrial short-chain enoyl-CoA hydratase, a key enzyme in fatty acid beta-oxidation and branched-chain amino acid degradation. The polyclonal population provides a heterogeneous knockout model suitable for studying loss-of-function effects without single-cell clonal selection.
The host 786-O cell line is derived from a primary clear cell renal cell carcinoma of a human male and is characterized by a VHL mutation leading to constitutive HIF activation and altered metabolic reprogramming. This VHL-deficient background makes 786-O a widely used model for investigating metabolic vulnerabilities in kidney cancer, particularly those related to lipid and amino acid utilization.
ECHS1 catalyzes the second step of mitochondrial fatty acid beta-oxidation, hydrating short-chain enoyl-CoA esters to L-3-hydroxyacyl-CoA. It also participates in the degradation of branched-chain amino acids isoleucine and valine, feeding into the TCA cycle via acetyl-CoA production. The enzyme functions within a multienzyme complex that includes ACADS (upstream) and HADH (downstream), with electron transfer via ETF. Its expression is regulated by PPARA and PPARGC1A in response to fasting/feeding signals. Knockout of ECHS1 disrupts these pathways, leading to accumulation of toxic intermediates, reduced ATP synthesis, and increased reactive oxygen species (ROS).
In the context of 786-O cells, ECHS1 loss further compromises mitochondrial metabolism, which is already rewired due to VHL deficiency. This combined dysregulation provides a potent model for investigating how ccRCC cells cope with impaired fatty acid oxidation and branched-chain amino acid catabolism. It also enables modeling of ECHS1 deficiency-related Leigh syndrome within a cancer metabolic framework, exploring the interplay between mitochondrial dysfunction and tumorigenesis.
This polyclonal knockout population is suitable for a range of applications, including studying mitochondrial fatty acid oxidation disorders, drug screening for mitochondrial therapies, and investigating metabolic reprogramming in renal cell carcinoma. Representative assays include Western blotting and RT-qPCR for metabolic enzyme expression, Seahorse metabolic flux analysis, targeted metabolomics (acylcarnitine profiling), enoyl-CoA hydratase activity measurement, and cellular viability assays under metabolic stress. For further information, please contact Ascent Research.
