The ECHS1 Knockout A2780 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A2780 human ovarian carcinoma cell line, featuring targeted disruption of the ECHS1 gene. This loss-of-function model enables investigation of mitochondrial short-chain enoyl-CoA hydratase deficiency and its consequences on fatty acid ??-oxidation and cellular energy metabolism. The polyclonal population retains genetic heterogeneity, reflecting a range of editing events, and is suitable for pooled functional studies without clonal selection.
The A2780 cell line originates from an untreated ovarian adenocarcinoma patient and is extensively characterized as a model for ovarian cancer biology, particularly for understanding cisplatin sensitivity and resistance mechanisms. As a human ovarian carcinoma line, A2780 cells exhibit robust mitochondrial oxidative metabolism and are well-suited for studying the intersection of oncogenic signaling and metabolic reprogramming. This background provides a clinically relevant context for dissecting the role of ECHS1 in cancer cell metabolism and therapeutic response.
ECHS1 encodes mitochondrial short-chain enoyl-CoA hydratase, which catalyzes the hydration of trans-2-enoyl-CoA to S-3-hydroxyacyl-CoA, the second step in the ??-oxidation cycle. Its activity is regulated by key metabolic sensors and transcription factors including PPARA, PPARGC1A, AMPK, SIRT1, and the hormones insulin and glucagon. ECHS1 functions downstream of fatty acid uptake and activation steps mediated by CPT1A and CPT2, and upstream of HADH and ACAA2, producing acetyl-CoA, NADH, and FADH2 that fuel the electron transport chain and ATP synthesis. Disruption of ECHS1 thus impairs short-chain fatty acid degradation, blunts acetyl-CoA and reducing equivalent generation, and triggers metabolic stress particularly under conditions of high lipid demand.
In A2780 ovarian cancer cells, defects in fatty acid oxidation can reveal metabolic vulnerabilities. ECHS1 loss reduces mitochondrial ??-oxidation capacity, potentially altering cellular NADH/NAD+ and ATP levels. Given the A2780 line??s utility in cisplatin studies, modulating fatty acid oxidation may intersect with mechanisms of drug sensitivity and resistance. Moreover, ECHS1 mutations are linked to Leigh syndrome and paroxysmal dyskinesia, making this polyclonal knockout model valuable for mitochondrial disease research, where metabolic adaptability and oxidative stress responses are critical.
Typical applications include Seahorse XF fatty acid oxidation assays to measure mitochondrial respiration in the presence of exogenous fatty acids, western blotting and RT-qPCR to confirm loss of ECHS1 and assess compensatory metabolic enzymes, and acylcarnitine profiling by mass spectrometry to detect accumulation of short-chain intermediates indicative of ??-oxidation blockade. Additional uses encompass cell viability assays under lipid-rich conditions, mitochondrial stress tests, and immunofluorescence for mitochondrial mass. This knockout model supports research into ovarian cancer metabolism, metabolic reprogramming, mitochondrial disease modeling, drug resistance studies, and redox biology. For further information, please contact Ascent Research.
