The EHHADH Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This genetically heterogeneous pool carries targeted disruption of the EHHADH gene, offering a robust loss-of-function model that preserves natural genetic variation within an epithelial context.
HT29 cells originate from a colorectal adenocarcinoma of a 44-year-old female and grow as adherent epithelial monolayers. They retain intestinal epithelial characteristics, including barrier function, mucus production, and ion transport, making them a relevant platform for studying colorectal cancer biology and intestinal metabolic processes.
EHHADH encodes the peroxisomal L-bifunctional protein, which catalyzes the second (enoyl-CoA hydratase) and third (3-hydroxyacyl-CoA dehydrogenase) steps of peroxisomal very long-chain fatty acid beta-oxidation. This enzyme is activated by PPARA and its ligands??fatty acids and fibrates??and functions downstream of the peroxisomal import receptor PEX5. Within the pathway, EHHADH interacts with ACOX1 and catalase, converting substrates from ABCD1-transported fatty acids into shorter-chain metabolites, ultimately yielding acetyl-CoA and hydrogen peroxide. Disruption of EHHADH leads to accumulation of very long-chain fatty acids and metabolic dysfunction.
In HT29 cells, EHHADH knockout impairs peroxisomal fatty acid metabolism, which can drive lipid accumulation and metabolic stress relevant to colorectal adenocarcinoma. This model is instrumental for exploring how peroxisomal dysfunction contributes to cancer cell homeostasis and for investigating the role of PPAR signaling in intestinal epithelial metabolism. It also provides a cellular system to mimic peroxisomal D-bifunctional protein deficiency in a colorectal context.
Typical applications include studying peroxisomal fatty acid oxidation in colorectal cancer, modeling peroxisomal disorders in intestinal epithelium, and examining metabolic reprogramming. Researchers can validate knockout efficiency via western blotting, RT-qPCR, and immunofluorescence, and assess functional consequences using fatty acid oxidation flux assays and very long-chain fatty acid profiling by LC-MS. Cell viability and proliferation assays further enable evaluation of EHHADH loss on tumor cell growth. For further details or to request a quote, please contact Ascent Research.
