The ECHDC1 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the NCI-H1299 human non-small cell lung adenocarcinoma cell line. This product provides a genetically disrupted ECHDC1 pool, eliminating the mitochondrial enoyl-CoA hydratase activity encoded by this gene.
The parental NCI-H1299 cell line was derived from a lymph node metastasis of a lung adenocarcinoma in a 43-year-old male and serves as a widely used epithelial model for non-small cell lung cancer (NSCLC) research. These cells retain key characteristics of metastatic lung cancer, making them especially suited for investigations into tumor metabolism.
ECHDC1 catalyzes the hydration of trans-2-enoyl-CoAs to 3-hydroxyacyl-CoAs in the mitochondrial fatty acid ??-oxidation pathway. Transcriptional regulation by PPAR?? and PPAR??, along with modulation by AMPK and insulin signaling, controls its expression. ECHDC1 operates in concert with other ??-oxidation enzymes and interacts with electron transfer flavoprotein (ETF) to couple fatty acid oxidation to the respiratory chain. Disruption of ECHDC1 impairs production of acetyl-CoA, NADH, and FADH2, leading to accumulation of enoyl-CoA intermediates and altered acylcarnitine profiles. This loss-of-function model forces a shift toward glycolytic metabolism, reflecting cancer-associated metabolic reprogramming.
In the NCI-H1299 NSCLC background, ECHDC1 knockout provides a unique opportunity to probe the reliance of lung adenocarcinoma cells on mitochondrial ??-oxidation. Many NSCLC cells depend on fatty acid oxidation for ATP production, redox homeostasis, and anabolic precursors, so ablation of this pathway can expose metabolic dependencies. The polyclonal population retains heterogeneity, allowing studies that capture a range of cellular responses to ??-oxidation disruption.
Representative assays include measurement of [U-13C]palmitate oxidation rates, Seahorse respirometry to quantify mitochondrial oxygen consumption, and LC-MS-based acylcarnitine profiling. Downstream analyses may examine acetyl-CoA levels, NADH/NAD+ ratios, and ATP synthesis under lipid-rich or glucose-limited conditions. This model can be integrated with modulators of PPARs, AMPK, or other ??-oxidation components to map signaling networks. Functional evaluations such as proliferation assays under varied lipid availability further characterize metabolic vulnerabilities. For more information, contact Ascent Research.
