The ECHS1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma line. This product provides a heterogeneous pool of cells with CRISPR-mediated disruption of the ECHS1 gene, enabling robust functional analyses without single-cell clonal bias. Suitable for pooled metabolic and functional genomic studies, these cells serve as a reliable system to investigate mitochondrial short-chain fatty acid oxidation and its role in cancer metabolism.
The parental NCI-H1975 cell line, a model of non-small cell lung cancer (NSCLC), originates from a female non-smoker and harbors wild-type EGFR and KRAS with a PIK3CA activating mutation. This genetic backdrop drives PI3K/AKT-mediated metabolic reprogramming, making it particularly relevant for studying lipid utilization and mitochondrial function in lung adenocarcinoma. ECHS1 knockout in these cells provides a tool to dissect the interplay between oncogenic signaling and fatty acid metabolism.
ECHS1 encodes mitochondrial short-chain enoyl-CoA hydratase, catalyzing the hydration of trans-2-enoyl-CoA to 3-hydroxyacyl-CoA in beta-oxidation. This step is integral to fatty acid degradation and the catabolism of valine, leucine, and isoleucine, yielding acetyl-CoA, NADH, and FADH2 for the TCA cycle. ECHS1 is regulated by metabolic sensors including PPARA, PPARGC1A, SIRT1, AMPK, and HNF4A, and interacts with enoyl-CoA substrates and mitochondrial matrix components. Within the beta-oxidation pathway, it collaborates with ACADS, HADH, ACAT1, and ACAA2. Disruption of ECHS1 impairs short-chain fatty acid catabolism, reducing acetyl-CoA pools and potentially shifting energy metabolism toward glycolysis.
In NCI-H1975 cells, ECHS1 knockout likely amplifies metabolic vulnerabilities imposed by PIK3CA-driven anabolic signaling. Loss of mitochondrial fatty acid oxidation may further shift the energy balance, impairing lipid-derived ATP production and increasing dependence on glucose. This model can reveal compensatory mechanisms, such as enhanced glycolysis or glutamine utilization, and serve as a platform to study ECHS1 deficiency??a disorder linked to Leigh syndrome, metabolic acidosis, and paroxysmal dyskinesia??within a cancer-relevant context.
Researchers can apply this knockout model in mitochondrial disease modeling, cancer metabolic reprogramming studies, and fatty acid oxidation analysis. Typical assays include Western blotting, RT-qPCR, Seahorse XF fatty acid oxidation stress tests, cellular ATP measurements, and LC-MS metabolomics for acyl-carnitine profiling. Additional assessments such as JC-1 mitochondrial membrane potential, glucose consumption/lactate production, cell proliferation/apoptosis, and mitochondrial morphology imaging can comprehensively characterize ECHS1 loss. For further details or custom options, contact Ascent Research.
