ECI2 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population from the NCI-H1975 human lung adenocarcinoma line, designed to disrupt the ECI2 gene encoding peroxisomal enoyl-CoA isomerase. The polyclonal format yields a diverse allele mixture, facilitating pooled loss-of-function analysis without clonal selection artifacts, and is ideal for studying peroxisomal ??-oxidation in EGFR-mutant cancer.
NCI-H1975 is an adherent epithelial cell line derived from a non-small cell lung adenocarcinoma, harboring the EGFR L858R/T790M double mutation that confers resistance to first-generation EGFR inhibitors while retaining sensitivity to third-generation agents like osimertinib. This well-characterized model supports research on EGFR-driven tumor biology, drug resistance, and metabolic reprogramming, and its stable growth properties enable high-throughput assays.
ECI2 (also known as PECI) encodes a peroxisomal enoyl-CoA isomerase that converts 3-cis to 2-trans enoyl-CoA intermediates, essential for continuing ??-oxidation of very long-chain and unsaturated fatty acids. The enzyme acts downstream of acyl-CoA oxidase 1 (ACOX1) and D-bifunctional protein (DBP), and cooperates with sterol carrier protein X (SCPx) and the ABCD1 transporter that imports substrates into peroxisomes. Transcription of ECI2 is controlled by PPAR?? and PPAR?? responding to fatty acids and nutritional cues. Through its catalytic action, ECI2 contributes to the production of acetyl-CoA and medium-chain acyl-CoAs, while diminishing cellular levels of very long-chain fatty acids, thus maintaining lipid homeostasis and preventing lipotoxicity.
In the NCI-H1975 background, ECI2 knockout likely disrupts peroxisomal lipid metabolism, potentially altering lipid droplet homeostasis, mitochondrial fatty acid oxidation, and membrane composition. Given the EGFR-mutant status and altered anabolic pathways, ECI2 loss may shift metabolic dependencies, sensitizing cells to lipid deprivation or oxidative stress. This model is valuable for dissecting how peroxisomal ??-oxidation interfaces with oncogenic signaling and for identifying metabolic vulnerabilities in lung adenocarcinoma.
Applications include functional analysis of peroxisomal ??-oxidation in EGFR-mutant lung cancer, metabolic flux studies with labeled fatty acids, lipidomics to quantify very long-chain fatty acid accumulation, and drug sensitivity profiling under lipid-modulated conditions. Validation assays such as western blotting and RT-qPCR confirm target disruption, while ATP and cell viability assays assess energetic and proliferative consequences. Fatty acid oxidation assays and lipidomic readouts provide detailed metabolic phenotyping. These polyclonal knockout cells thus offer a robust system to investigate the crosstalk between peroxisomal function and tumor fitness. For further inquiries, please contact Ascent Research.
