The ECI1 Knockout NCI-H1299 Polyclonal Cells product is a CRISPR/Cas9-mediated polyclonal knockout cell population targeting the ECI1 gene in the human non-small cell lung carcinoma cell line NCI-H1299. This polyclonal knockout pool, generated by transient delivery of ribonucleoprotein complexes, offers a heterogeneous mixture of edited alleles, enabling robust loss-of-function studies without clonal isolation. The product is designed for researchers investigating fatty acid metabolism and its intersection with cancer biology, providing a physiologically relevant model system to dissect mitochondrial beta-oxidation pathways.
NCI-H1299 cells are derived from a lymph node metastasis of a patient with non-small cell lung cancer, serving as a widely utilized model for metastatic lung cancer. These adherent epithelial cells harbor characteristics that recapitulate the aggressive behavior of advanced lung tumors, including altered metabolic dependencies. Their ability to form tumors in xenograft models and their well-characterized genomic landscape make them suitable for studying the metabolic adaptations that drive tumor progression and metastasis, particularly in the context of lipid utilization.
ECI1 encodes mitochondrial enoyl-CoA delta isomerase, a critical enzyme in the beta-oxidation of unsaturated fatty acids. This enzyme catalyzes the isomerization of cis-3-enoyl-CoA to trans-2-enoyl-CoA, enabling flux through the fatty acid degradation pathway. ECI1 is transcriptionally regulated by upstream factors such as PPAR?? and PPAR??, which respond to insulin/glucagon signaling and nutritional status. It functions within a multienzyme complex that includes interacting partners like the trifunctional protein (HADHA/HADHB) and electron transfer flavoprotein (ETF), and operates sequentially with enzymes such as enoyl-CoA hydratase (ECHS1) and downstream targets like ACADVL, HADH, and ACAA2. Disruption of ECI1 impairs the conversion of unsaturated fatty acyl-CoAs, leading to reduced acetyl-CoA production, diminished mitochondrial beta-oxidation flux, and potential alterations in lipid accumulation and energy homeostasis.
In the NCI-H1299 lung cancer background, ECI1 knockout provides a powerful tool to interrogate the role of fatty acid oxidation in cancer metabolism. Lung cancer cells often rewire their metabolic networks to support proliferation and survival under stress; this knockout model enables dissection of how impaired unsaturated fatty acid degradation affects metabolic reprogramming, mitochondrial function, and tumorigenic properties. Given the frequent dysregulation of lipid metabolism in non-small cell lung cancer, this model is particularly relevant for exploring vulnerabilities that may arise from compromised beta-oxidation, potentially informing therapeutic strategies that target metabolic pathways.
This knockout polyclonal pool is suited for a range of experimental applications, including fatty acid oxidation assays using radiolabeled substrates, Seahorse metabolic flux analysis to measure oxygen consumption rates, and lipid accumulation staining to visualize neutral lipid droplets. Researchers can employ Western blotting and RT-qPCR to confirm ECI1 disruption, and perform cell proliferation and apoptosis assays to assess functional consequences. It also facilitates drug sensitivity studies targeting metabolic enzymes like CPT1A or CPT2. For further information, please contact Ascent Research.
