ECHDC1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma cell line. This heterogeneous pool contains cells with targeted disruption of the ECHDC1 gene, creating a loss-of-function model for investigating mitochondrial fatty acid oxidation and related metabolic pathways. The polyclonal format avoids clonal selection, offering a population-level representation of knockout phenotypes for robust functional studies.
The NCI-H1975 cell line originates from a non-small cell lung cancer (NSCLC) patient and harbors well-characterized EGFR mutations (L858R and T790M) along with a PIK3CA mutation. This genetic background is widely used to study EGFR-targeted therapy resistance and metabolic reprogramming in lung adenocarcinoma. The co-occurrence of these oncogenic drivers alters cellular metabolism, providing a relevant context to explore how ECHDC1 loss influences cancer cell bioenergetics and signaling.
ECHDC1 encodes a mitochondrial enoyl-CoA hydratase/decarboxylase that converts ethylmalonyl-CoA to butyryl-CoA, a key step in odd-chain fatty acid and branched-chain amino acid catabolism. It is regulated by PPAR?? and PGC-1?? transcriptional programs and AMPK signaling. ECHDC1 interacts with acyl-CoA dehydrogenases and electron transfer flavoprotein (ETF), and its product butyryl-CoA enters ketone body synthesis, the TCA cycle, and histone butyrylation. Loss of function causes ethylmalonic acid accumulation and metabolic acidosis, modeling features of ethylmalonic encephalopathy.
In NCI-H1975 cells, ECHDC1 knockout provides a model to investigate the interplay between oncogenic signaling and mitochondrial metabolism. The EGFR and PIK3CA mutations drive anabolic growth and may confer dependency on fatty acid oxidation and branched-chain amino acid pathways. This knockout enables studies of metabolic vulnerability in lung cancer, potential synthetic lethality, and ethylmalonic encephalopathy-like defects within a cancer cell context, aiding research into mitochondrial dysfunction and metabolic reprogramming.
Assays with these cells include Western blotting and RT-qPCR for knockout confirmation, Seahorse metabolic flux analysis, fatty acid oxidation measurement, and LC-MS metabolite profiling to detect ethylmalonic acid and TCA cycle shifts. Functional readouts such as cell viability, migration, and invasion assays, along with immunofluorescence for mitochondrial markers, support dissection of ECHDC1’s role in cancer phenotypes. These applications suit metabolic enzyme inhibitor screening and mechanistic studies. For product inquiries, please contact Ascent Research.
