The ECI1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human lung adenocarcinoma cell line NCI-H1975 (Homo sapiens). This product provides a heterogeneous pool of cells bearing targeted gene disruption of ECI1 (enoyl-CoA delta isomerase 1), generated without clonal isolation, thus preserving the natural diversity of editing outcomes. The polyclonal format offers a representative loss-of-function model for investigating ECI1-dependent metabolic processes while avoiding potential artifacts of clonal selection.
The host NCI-H1975 line is an epithelial cell model of non-small cell lung adenocarcinoma carrying activating EGFR L858R/T790M mutations and a TP53 tumor suppressor mutation. This clinically relevant genetic background recapitulates key aspects of lung cancer biology, including aberrant growth signaling and genomic instability, making it particularly suitable for studying the interplay between oncogenic drivers and metabolic reprogramming.
ECI1 encodes a mitochondrial enzyme catalyzing the isomerization of 3-cis-enoyl-CoA to 3-trans-enoyl-CoA, a required step in unsaturated fatty acid ??-oxidation. This enzyme operates within a pathway that includes ACSL, CPT1, ACADM, HADHA, HADHB, and ACAA2, ultimately producing acetyl-CoA, TCA cycle intermediates, and ATP. ECI1 interacts with HADHA, HADHB, and ACADVL within the mitochondrial trifunctional protein complex. Its expression is regulated by PPARA and PPARD, which are activated by AMPK. Disruption of ECI1 by CRISPR/Cas9 blocks isomerization, leading to enoyl-CoA accumulation and reduced energy output.
In NCI-H1975 lung adenocarcinoma cells, ECI1 disruption impairs unsaturated fatty acid ??-oxidation, leading to reduced acetyl-CoA and ATP. The compromised energy metabolism may be particularly consequential in the p53-mutant context, sensitizing cells to metabolic stress. This knockout model facilitates investigation of fatty acid oxidation dependency in EGFR-driven lung cancer and the identification of synthetic lethal interactions.
Applications include metabolic flux analysis (Seahorse), 14C-palmitate oxidation assays, ATP and MTT viability measurements, and pathway validation by western blotting and RT-qPCR. Metabolomic profiling can detect enoyl-CoA intermediate accumulation. This model is suitable for drug screening targeting fatty acid metabolism and for modeling ECI1 deficiency. For further details, contact Ascent Research.
