The GRHPR Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human hepatic adenocarcinoma cell line. This heterogeneous mixture of GRHPR-disrupted cells provides a robust loss-of-function model that avoids clonal selection artifacts, making it ideal for population-level metabolic studies and pooled genetic screens. By eliminating glyoxylate reductase/hydroxypyruvate reductase activity, the model allows systematic investigation of hepatic glyoxylate metabolism and oxalate homeostasis in a liver-relevant context.
SK-HEP-1 is a well-established cell line originating from the ascites of a patient with hepatic adenocarcinoma, and it displays both epithelial and endothelial characteristics. Widely used as a hepatocellular carcinoma model, these cells retain hepatocyte-like functions including gluconeogenesis, lipid metabolism, and detoxification pathways. Their adherent growth and robust culturability facilitate a broad range of in vitro assays, making them an appropriate host for exploring the consequences of GRHPR loss in a liver-derived system.
GRHPR encodes glyoxylate reductase/hydroxypyruvate reductase, an NADPH-dependent oxidoreductase that catalyzes the reduction of glyoxylate to glycolate and hydroxypyruvate to D-glycerate. This reaction is essential for preventing the accumulation of oxalate, a nephrotoxic metabolite. GRHPR activity is putatively regulated by hepatocyte nuclear factors (HNFs) and metabolic signals, and it requires NADPH as a cofactor. The enzyme operates at a critical junction of the glyoxylate and dicarboxylate metabolism pathway, interacting functionally with alanine-glyoxylate aminotransferase (AGT) and lactate dehydrogenase (LDH). Disruption of GRHPR in SK-HEP-1 cells impairs glyoxylate clearance, resulting in oxalate buildup that models the metabolic defect observed in primary hyperoxaluria type 2.
In the SK-HEP-1 hepatic environment, GRHPR knockout provides a physiologically relevant platform to dissect liver-specific mechanisms of oxalate homeostasis and the pathogenesis of hyperoxaluria type 2. Since the liver is the primary site of glyoxylate detoxification, these cells express the requisite transporters and enzymes for glyoxylate handling. The knockout enables exploration of oxalate-induced cytotoxicity, its impact on hepatocyte function, and the interplay with serine biosynthesis. Furthermore, conditioned media from these cells can be used to study paracrine effects on renal epithelial cells, mimicking the liver-kidney axis in oxalate nephropathy.
This polyclonal knockout product supports diverse applications, including disease modeling of primary hyperoxaluria type 2, drug screening for oxalate-lowering therapeutics, and mechanistic studies of hepatic metabolism. Representative assays include NADPH oxidation enzyme assays to verify GRHPR inactivation, LC-MS-based oxalate and glycolate quantification, cell viability tests under oxalate challenge, and transcriptomic profiling via RNA-seq. Co-culture systems can investigate hepatorenal communication. For further technical information or to discuss custom gene-editing services, please contact Ascent Research.
