The ECHDC3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from human HT29 colorectal adenocarcinoma cells, featuring disruption of the ECHDC3 gene. This polyclonal population comprises a heterogeneous pool of gene-edited alleles generated by CRISPR/Cas9-mediated target-gene disruption, offering a robust loss-of-function model for studying ECHDC3 function. Supplied as a live cell stock, it is ready for immediate culture and experimental use.
The parental HT29 cell line is a well-established model of human colorectal adenocarcinoma with epithelial morphology. Widely used for studying intestinal epithelial biology and colorectal cancer, HT29 cells form polarized monolayers and respond to microenvironmental signals, providing a pathophysiologically relevant context for dissecting tumor cell metabolism. Their epithelial characteristics make them ideal for investigating mitochondrial lipid handling in a colonic tumor setting. HT29 cells are frequently employed to model colorectal cancer progression and drug response.
ECHDC3 encodes a mitochondrial enoyl-CoA hydratase that catalyzes hydration of trans-2-enoyl-CoA to 3-hydroxyacyl-CoA in the fatty acid beta-oxidation spiral. It operates within a multienzyme complex interacting with HADHA, HADHB, ACADVL, and ECHS1. ECHDC3 expression is regulated by PPARGC1A, PPARA, MLXIPL, and SREBF1, which orchestrate lipid catabolic programs. Downstream, ECHDC3 activity impacts mitochondrial oxidative capacity, ATP production, and reactive oxygen species levels, linking fatty acid oxidation to cellular energy homeostasis.
In HT29 colorectal adenocarcinoma cells, ECHDC3 knockout is predicted to impair mitochondrial fatty acid oxidation, potentially causing metabolic reprogramming toward glycolysis or glutaminolysis. This may alter lipid accumulation, growth under lipid-rich conditions, and sensitivity to metabolic stress. Given the metabolic dysregulation common in colorectal cancers, this model is valuable for studying how disrupted mitochondrial lipid metabolism influences tumor phenotypes such as proliferation, survival, and drug resistance.
These polyclonal knockout cells are suitable for lipid metabolism studies, cancer metabolism research, and metabolic drug screening. Researchers can confirm ECHDC3 disruption via Western blotting and RT-qPCR, and evaluate functional impacts using fatty acid oxidation assays, Seahorse-based mitochondrial respiration, Oil Red O lipid staining, and viability assays under lipid-rich or nutrient-stressed conditions. They also enable drug sensitivity profiling to identify compounds targeting metabolic vulnerabilities. For additional information or custom engineering inquiries, please contact Ascent Research.
