The H6PD knockout HT29 polyclonal cells are a CRISPR/Cas9-edited cell population generated from the human colorectal adenocarcinoma cell line HT29. This product provides a loss-of-function model for hexose-6-phosphate dehydrogenase (H6PD) through targeted gene disruption, enabling investigation of its role in endoplasmic reticulum (ER) lumenal NADPH generation and glucocorticoid metabolism. As a polyclonal pool, the cells retain genomic diversity at the targeted locus while collectively representing the gene knockout phenotype, making them suitable for population-level functional assays.
The HT29 cell line is a well-characterized human colorectal adenocarcinoma line with epithelial morphology, originally derived from a primary tumor. These cells have been extensively employed in studies of intestinal epithelial differentiation, transport processes, and colorectal cancer biology. Their ability to form polarized monolayers and produce mucin makes them particularly valuable for examining epithelial barrier function and tissue-specific metabolic pathways in vitro.
H6PD encodes an ER-resident enzyme that oxidizes glucose-6-phosphate to generate NADPH within the lumen. This NADPH pool is an obligate cofactor for 11??-hydroxysteroid dehydrogenase type 1 (HSD11B1), which catalyzes the conversion of inactive cortisone to active cortisol. Substrate supply relies on the glucose-6-phosphate transporter SLC37A4, and H6PD interacts with ER chaperones to maintain functional integrity. Upstream, H6PD expression is regulated by PPAR?? and induced by ER stress, while downstream it modulates NADPH-dependent ER reductases and cortisol production. Consequently, H6PD serves as a critical node linking the pentose phosphate pathway to local glucocorticoid activation and redox homeostasis.
In the HT29 colorectal cancer background, disruption of H6PD is expected to impair local cortisol regeneration, thereby altering glucocorticoid-responsive gene expression that influences cell proliferation, differentiation, and apoptosis. Given the colon??s exposure to circulating glucocorticoids, H6PD-dependent cortisol production may also affect epithelial barrier integrity and inflammatory signaling. Moreover, loss of luminal NADPH generation disrupts ER redox balance, potentially exacerbating ER stress??a hallmark of cancer cell metabolism. This model thus offers a physiologically relevant system to dissect the intersection of glucocorticoid metabolism and redox control in colorectal tumorigenesis.
Researchers can utilize these knockout cells to investigate glucocorticoid metabolism in colorectal cancer, ER stress, and metabolic reprogramming. Representative assays include cortisol ELISA, NADPH/NADP+ ratio determinations, Western blotting for ER stress markers, transwell epithelial barrier assays, and cell viability analyses. The polyclonal format is ideal for population-based experiments prioritizing phenotypic robustness. For further details, pricing, or technical support, please contact Ascent Research.
