The ECHDC3 Knockout NCI-H1299 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the ECHDC3 gene has been disrupted via CRISPR/Cas9-mediated gene disruption. This product provides a heterogeneous pool of NCI-H1299 cells carrying targeted loss-of-function mutations in ECHDC3, generating a versatile model for studying mitochondrial fatty acid ??-oxidation in a metastatic non-small cell lung carcinoma (NSCLC) background. The polyclonal format avoids single-cell cloning artifacts and preserves population-level genetic heterogeneity, making it suitable for metabolic flux analyses and pooled screening applications.
The parental NCI-H1299 cell line is a human lung carcinoma line derived from lymph node metastasis of a large cell carcinoma, widely utilized as a model for metastatic NSCLC. This p53-deficient line exhibits aggressive growth phenotypes and altered metabolic dependencies, including reliance on fatty acid oxidation for energy production and survival under nutrient stress. Its origins from a metastatic site make it particularly relevant for investigating metabolic reprogramming associated with tumor dissemination.
ECHDC3 encodes a mitochondrial enoyl-CoA hydratase that catalyzes the second step of the fatty acid ??-oxidation cycle, converting enoyl-CoA thioesters to 3-hydroxyacyl-CoA intermediates. This enzyme functions within the mitochondrial ??-oxidation multienzyme complex, interacting closely with HADHA, HADHB, and ECHS1 to complete long-chain fatty acid degradation. Upstream, ECHDC3 expression is regulated by the PPARA?CPPARGC1A (PGC-1??)?CNRF1 transcriptional axis, which coordinates mitochondrial biogenesis and lipid catabolism. Downstream, its activity contributes to acetyl-CoA and ATP production, feeding the TCA cycle and cellular energy homeostasis. Disruption of ECHDC3 thus uncouples fatty acid substrate supply from mitochondrial oxidative metabolism.
In NCI-H1299 cells, fatty acid oxidation is a critical metabolic pathway supporting growth, survival, and metastatic potential. By disrupting ECHDC3, this knockout model impairs the ??-oxidation flux, forcing cells to rely on alternative carbon sources or to undergo metabolic adaptation. This is particularly insightful for studying how NSCLC cells respond to bioenergetic stress imposed by fatty acid oxidation inhibition, a vulnerability observed in tumors with high PPARGC1A activity. The model can be used to dissect crosstalk between lipid metabolism and oncogenic signaling in a p53-null background, and to identify compensatory mechanisms that may limit therapeutic efficacy of FAO inhibitors such as etomoxir.
Typical experimental applications include metabolic flux analysis using palmitate-driven oxygen consumption measurements, metabolomics profiling via LC-MS to track TCA cycle intermediates, and western blot assessment of ECHDC3 and interacting ??-oxidation enzymes. This polyclonal pool is also amenable to drug sensitivity screening under nutrient-restricted conditions, invasion/migration assays to probe the role of lipid metabolism in metastasis, and co-culture experiments modeling tumor microenvironment interactions. For further technical details, customized project support, or additional validation information, please contact Ascent Research.
