The GRHPR Knockout HEK293T Polyclonal Cells consist of a pool of HEK293T cells subjected to CRISPR/Cas9-mediated gene disruption targeting the GRHPR locus, generating a heterogeneous polyclonal knockout population. This product provides a versatile loss-of-function model for investigating glyoxylate metabolism and oxalate-related diseases without the bias of single-clone selection. The polyclonal format retains genetic diversity, enabling robust and reproducible functional studies in a well-characterized cellular background.
HEK293T cells are a human embryonic kidney epithelial cell line immortalized by stable expression of the SV40 large T antigen. This line is widely utilized for its high transfection efficiency, rapid growth, and capacity for high-level protein production. Deriving from renal epithelium, HEK293T cells maintain relevant metabolic pathways and signaling mechanisms inherent to kidney cells, making them an appropriate system for studying renal metabolism, including glyoxylate and oxalate handling.
GRHPR encodes glyoxylate reductase/hydroxypyruvate reductase, an enzyme that catalyzes the NADPH-dependent reduction of glyoxylate to glycolate and hydroxypyruvate to D-glycerate. This reaction is central to glyoxylate detoxification and serine metabolism. GRHPR expression is regulated by PPARGC1A, insulin, and glucagon, linking metabolic hormone signaling to oxalate homeostasis. By consuming glyoxylate, GRHPR limits its oxidation to oxalate, a reaction mediated by lactate dehydrogenase. Consequently, GRHPR acts upstream of oxalate production, and its dysfunction leads to glyoxylate accumulation and enhanced oxalate formation, promoting calcium oxalate precipitation. The balance between GRHPR and alanine-glyoxylate aminotransferase further determines the metabolic fate of glyoxylate.
In the HEK293T background, GRHPR knockout recapitulates the metabolic defect of primary hyperaluria type 2, a rare disorder characterized by recurrent calcium oxalate nephrolithiasis and renal failure. These polyclonal knockout cells exhibit impaired glyoxylate reduction, leading to elevated intracellular oxalate levels and increased sensitivity to oxalate-induced cytotoxicity. As kidney epithelial cells, they provide a physiologically relevant platform to study oxalate handling, crystal adhesion, and epithelial injury responses. This model can be used to dissect the molecular mechanisms of oxalate toxicity and to identify targets for therapeutic intervention.
This knockout product is suited for a range of experimental applications, including enzyme activity assays measuring NADPH consumption, western blotting and RT-qPCR for expression analysis, intracellular oxalate quantification, and cell viability assays under oxalate stress. Researchers can employ this system to study metabolic flux through glyoxylate pathways, screen for oxalate-lowering agents, and model kidney stone pathology in vitro. For further technical details or to discuss custom modifications, please contact Ascent Research.
