The ECI1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line. This product provides a genetically heterogeneous pool of cells harboring disruptions in the ECI1 gene, enabling loss-of-function studies in a physiologically relevant epithelial context. The polyclonal format avoids clonal selection artifacts and maintains biological variability, making it suitable for pooled screening and bulk metabolic assays.
The HT29 cell line is a well-established model of human intestinal epithelial biology, originally isolated from a primary colon adenocarcinoma. HT29 cells retain many features of differentiated enterocytes and are widely employed to investigate colorectal cancer pathophysiology, cellular metabolism, and epithelial barrier function. They exhibit robust growth and are amenable to genetic manipulation, providing a consistent platform for studying gene function in colorectal cancer.
ECI1 encodes enoyl-CoA delta isomerase 1, a mitochondrial enzyme that isomerizes 3-cis and 3-trans enoyl-CoA esters to 2-trans-enoyl-CoA, a critical step in the beta-oxidation of unsaturated fatty acids. ECI1 functions within the mitochondrial fatty acid oxidation pathway alongside acyl-CoA dehydrogenase, enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and ketoacyl-CoA thiolase. Its activity is transcriptionally regulated by PPAR??, PPAR??, and HNF4A, and it interacts with the mitochondrial trifunctional protein complex, electron transfer flavoprotein (ETF), and ETF-dehydrogenase to facilitate efficient energy production from unsaturated fatty acids. Disruption of ECI1 leads to accumulation of unsaturated enoyl-CoA intermediates and reduces generation of acetyl-CoA, impairing mitochondrial energy metabolism.
In HT29 colorectal cancer cells, ECI1 knockout is expected to impair the utilization of unsaturated fatty acids for energy production, potentially shifting metabolic reliance toward glycolysis or saturated fatty acid oxidation. This alteration in lipid homeostasis may impact cell proliferation, survival, and response to metabolic stress, making the knockout model a valuable tool for dissecting the role of fatty acid metabolism in colorectal tumorigenesis. Studies using these cells can reveal adaptive metabolic reprogramming and identify vulnerabilities exploitable for therapeutic intervention.
Research applications of the ECI1 Knockout HT29 Polyclonal Cells include metabolic flux analysis using isotopically labeled fatty acids, fatty acid oxidation assays, cell proliferation and colony formation assays, ATP measurement, and lipid droplet staining. The polyclonal population is suitable for bulk biochemical assays, western blotting of key metabolic enzymes such as ETF-dehydrogenase and members of the trifunctional protein complex, and transcriptomic analyses via RT-qPCR or RNA-seq to assess changes in PPAR?? and PPAR?? target gene expression. This model supports functional validation of ECI1 in colorectal cancer metabolism, lipid metabolism disorder studies, and drug target validation. For more information and expert support, please contact Ascent Research.
