The HSD17B4 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the HSD17B4 gene in the human HeLa cell background. This loss-of-function model enables investigation of peroxisomal fatty acid ??-oxidation and steroid metabolism without the need for single-cell clonal selection. The targeted gene disruption abolishes the enzymatic activity of the HSD17B4-encoded multifunctional protein, providing a flexible tool for studying metabolic pathway perturbations in a cancer cell context.
HeLa cells are an immortalized epithelial cell line derived from cervical adenocarcinoma, widely used for cancer biology and general cell research due to their robust proliferation and ease of manipulation. They provide a consistent and scalable system for genetic perturbation, facilitating reproducible metabolic and signaling studies. Their well-characterized biology and established protocols make them suitable for investigating peroxisomal function and cancer metabolism.
HSD17B4 encodes a peroxisomal enzyme with 17??-hydroxysteroid dehydrogenase activity that catalyzes the second and third steps of peroxisomal ??-oxidation of very long-chain fatty acids and participates in steroid hormone metabolism. It is transcriptionally regulated by PPAR?? in response to nutritional status and functions in a pathway alongside ACOX1, SCPx, and 3-ketoacyl-CoA thiolase. The enzyme interacts with peroxisomal matrix import machinery and is essential for converting enoyl-CoA esters into 3-ketoacyl-CoA intermediates, linking fatty acid degradation to bile acid synthesis and steroid metabolism.
In the HeLa cell context, HSD17B4 knockout disrupts peroxisomal ??-oxidation, leading to potential accumulation of very long-chain fatty acids and altered C27 bile acid intermediate production. This model is particularly relevant for studying peroxisomal disorders, D-bifunctional protein deficiency, and the metabolic reprogramming of cancer cells, where lipid metabolism plays a critical role. The loss of HSD17B4 may sensitize cells to lipotoxicity or oxidative stress, providing a platform for exploring therapeutic interventions in peroxisomal dysfunction and cancer metabolism.
Researchers can employ HSD17B4 Knockout HeLa Polyclonal Cells in a variety of assays, including very long-chain fatty acid profiling by GC-MS, peroxisomal ??-oxidation activity measurements, Western blotting for HSD17B4 protein, RT-qPCR for mRNA quantification, immunofluorescence to assess peroxisome integrity, and metabolic flux analysis to map lipid utilization. These applications support studies in peroxisomal disorder modeling, fatty acid metabolism, steroid hormone biosynthesis, and cancer cell metabolic adaptation. For further details or to inquire about custom solutions, please contact Ascent Research.
