The ECI2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HT29 human colorectal adenocarcinoma cells, with targeted disruption of the ECI2 gene. This polyclonal format provides a genetically heterogeneous pool of cells, minimizing clonal artifacts and representing a population-level knockout response. The cells maintain the background characteristics of HT29 while eliminating ECI2 function.
The HT29 cell line originates from a 44-year-old female patient with colorectal adenocarcinoma and serves as a widely used model of intestinal epithelium. These adherent epithelial cells carry mutations in APC and TP53 but remain KRAS wild-type, reflecting a common colorectal cancer mutational profile. This genetic background makes HT29 particularly suitable for studying tumor metabolism and signaling.
ECI2 encodes a mitochondrial enoyl-CoA isomerase essential for ??-oxidation of unsaturated fatty acids by isomerizing 3-cis and 3-trans enoyl-CoA intermediates to enable their entry into the core ??-oxidation spiral. Expression of ECI2 is regulated by PPAR?? and PGC-1?? in response to nutritional status. The enzyme interacts with the trifunctional protein subunits HADHA and HADHB and long-chain acyl-CoA dehydrogenase (ACADL) within a pathway that includes CPT1A, ACADVL, and ACAA2. Disruption of ECI2 impairs mitochondrial utilization of unsaturated fatty acids, potentially leading to a shift toward glycolytic metabolism or lipid storage.
In the context of HT29 colorectal cancer cells, loss of ECI2 creates a model to investigate metabolic reprogramming driven by defective unsaturated fatty acid oxidation. The coexisting APC and TP53 mutations may synergize with ECI2 deficiency to reveal metabolic vulnerabilities, such as enhanced dependence on glycolysis or altered lipid droplet dynamics. This system allows examination of how cancer cells adapt to impaired lipid catabolism and the role of nutritional cues.
This knockout polyclonal cell pool is optimized for cancer metabolism research, including studies on fatty acid oxidation in colorectal cancer, metabolic reprogramming, and lipid metabolism disorders. Applications include Seahorse metabolic flux analysis to measure fatty acid oxidation-dependent oxygen consumption, fatty acid oxidation assays using labeled substrates (e.g., [U-13C]oleate), Oil Red O staining for lipid droplets, and cell proliferation and migration assays. Molecular analyses can assess PPAR??, PGC-1??, HADHA, and HADHB expression by RT-qPCR and Western blotting. For technical support and additional information, contact Ascent Research.
