The H6PD Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the H6PD gene in the near-haploid HAP1 cell line. This product provides a genetically defined loss-of-function model for investigating the microsomal glucose-6-phosphate dehydrogenase (H6PD) enzyme and its role in endoplasmic reticulum (ER) NADPH generation and glucocorticoid metabolism.
The HAP1 cell line is a human near-haploid fibroblast-like line originally derived from the KBM-7 chronic myeloid leukemia cell line. Its near-haploid karyotype simplifies genetic manipulation and functional genomics studies, making it a widely used platform for knockout screens and loss-of-function analyses. The HAP1 background enables efficient CRISPR/Cas9 editing and largely avoids the confounding effects of diploid genetic redundancy, facilitating clear genotype?Cphenotype correlations in cellular assays.
H6PD encodes a microsomal glucose-6-phosphate dehydrogenase that catalyzes the conversion of glucose-6-phosphate to 6-phosphogluconolactone, generating NADPH exclusively within the ER lumen. This NADPH pool serves as an essential cofactor for 11??-hydroxysteroid dehydrogenase type 1 (HSD11B1), which reduces inactive cortisone to active cortisol, thereby modulating local glucocorticoid receptor signaling. H6PD is transcriptionally regulated by PPARG, insulin, and glucocorticoid receptor pathways. Its activity is functionally coupled to the glucose-6-phosphate transporter SLC37A4 (G6PT) and interacts with ER oxidoreductases to maintain redox homeostasis. Disruption of H6PD abolishes NADPH supply to HSD11B1, impairing cortisone-to-cortisol conversion and altering downstream glucocorticoid-responsive gene expression.
In the HAP1 haploid background, knockout of H6PD creates a clean null-mutant system for interrogating the cortisone?Ccortisol shuttle and ER redox biology without interference from a second functional allele. This model is particularly valuable for dissecting the metabolic interplay between the pentose phosphate pathway and glucocorticoid activation, as well as for identifying genetic or pharmacological modifiers of HSD11B1 activity. The near-haploid format also facilitates unbiased forward genetic screens and synthetic lethal interaction studies in pathways dependent on ER NADPH generation.
Typical research applications include functional studies of cortisone reductase deficiency, investigation of glucocorticoid-driven adipocyte differentiation, and exploration of insulin resistance mechanisms linked to hepatic and adipose tissue cortisol tone. The cells can be employed in downstream assays such as Western blotting for H6PD and HSD11B1 expression, enzymatic measurement of NADPH/NADP+ ratios, cortisone-to-cortisol conversion assays using LC-MS, and RT-qPCR analysis of glucocorticoid-responsive genes like PEPCK and G6PC. Furthermore, the H6PD knockout HAP1 model serves as a robust platform for screening HSD11B1 inhibitors and for assessing crosstalk between insulin signaling and ER redox regulation. For additional information or to discuss custom gene-edited cell models, please contact Ascent Research.
