The HSD17B4 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the HSD17B4 gene in the HEK293T host cell background. This product provides a loss-of-function model to study the roles of the peroxisomal multifunctional enzyme HSD17B4 in steroid hormone metabolism and fatty acid oxidation. The polyclonal nature ensures a heterogeneous knockout population, suitable for experiments where diverse genetic perturbations model biological variability without requiring single-cell clonal isolation.
The HEK293T cell line is derived from human embryonic kidney epithelial cells and stably expresses the SV40 large T antigen, which enhances episomal replication of plasmids and allows high-level protein expression. These adherent cells are widely employed in mammalian expression systems and gene editing applications due to their robust growth and transfection efficiency. The HEK293T background provides a well-characterized platform for investigating gene function, particularly in pathways related to metabolism and signal transduction.
HSD17B4 encodes a peroxisomal enzyme that catalyzes the oxidation of estradiol to estrone and participates in the beta-oxidation of very long-chain fatty acids (VLCFAs). It is a key component of peroxisomal lipid metabolism and steroid hormone biosynthesis. The enzyme??s activity is regulated by upstream factors including the nuclear receptor PPAR??, thyroid hormone, retinoic acid, and the peroxin PEX19. HSD17B4 functions downstream of PPAR???CRXR signaling, driving fatty acid oxidation to generate acetyl-CoA, and directly converts estradiol to estrone, thereby modulating estrogen signaling pathways. It interacts with peroxisomal proteins such as SCP2, PEX5, ACOX1, and the DBP complex, forming an integrated metabolic network.
Knockout of HSD17B4 in HEK293T cells eliminates the oxidation of estradiol and the degradation of VLCFAs, creating a cellular model that mimics aspects of peroxisomal biogenesis disorders. This model is particularly relevant for studying D-bifunctional protein deficiency, Perrault syndrome, Zellweger spectrum disorders, and adrenoleukodystrophy-like conditions. By removing HSD17B4 activity, researchers can dissect the enzyme??s contribution to bile acid biosynthesis, peroxisomal lipid metabolism, and steroid hormone homeostasis, using a human cell context that is amenable to genetic manipulation and high-throughput screening.
The HSD17B4 Knockout HEK293T Polyclonal Cells are suitable for a variety of research applications, including mechanistic studies of estrogen signaling, drug metabolism screening, and peroxisomal disorder modeling. Representative assays with this model include LC-MS/MS quantification of estradiol/estrone ratios, radiolabeled VLCFA beta-oxidation assays, immunofluorescence staining for peroxisomal markers, Western blotting and RT-qPCR for HSD17B4 expression validation, and VLCFA accumulation measurements. Researchers can also analyze bile acid intermediates to investigate downstream metabolic effects. For additional information or custom inquiries, please contact Ascent Research.
