GRHPR Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 colorectal adenocarcinoma line, with targeted disruption of the GRHPR gene. This loss-of-function model enables robust investigation of glyoxylate reductase/hydroxypyruvate reductase in an intestinal epithelial background. The polyclonal format avoids single-cell clone biases and is well-suited for pooled screening and population-level assays.
HT29 is a widely characterized human cell line established from a female colorectal adenocarcinoma. It exhibits enterocyte-like differentiation, forming polarized monolayers with brush border enzymes, and serves as a standard model for intestinal drug absorption, nutrient transport, and colorectal cancer research. Its reproducible culture properties and genetic stability provide a reliable host for CRISPR-edited derivatives.
GRHPR encodes a cytosolic NADPH-dependent homodimeric enzyme that reduces glyoxylate to glycolate and hydroxypyruvate to D-glycerate, representing a pivotal control point in glyoxylate detoxification and serine metabolism. The enzyme interacts metabolically with glycolate oxidase (GO), alanine-glyoxylate aminotransferase (AGT), hydroxyacid oxidase 1 (HAO1), and lactate dehydrogenase (LDH) to govern oxalate production. In serine biosynthesis, GRHPR connects with phosphoglycerate dehydrogenase (PHGDH) and serine hydroxymethyltransferase (SHMT), integrating hydroxypyruvate reduction with one-carbon flux.
Knockout of GRHPR in HT29 cells abolishes enzyme activity, causing glyoxylate and hydroxypyruvate accumulation, elevated oxalate synthesis, and disrupted serine metabolism. This phenotype mirrors primary hyperoxaluria type 2 defects and induces oxidative stress with altered proliferation, revealing metabolic liabilities relevant to colorectal cancer. The colonic epithelial context further enables studies of intestinal oxalate transport and its systemic impact on urolithiasis.
Applications include mechanistic dissection of glyoxylate and dicarboxylate metabolism, disease modeling of primary hyperoxaluria type 2, and cancer metabolism research. The model is validated via Western blot, RT-qPCR, and glyoxylate reductase activity assays, with metabolic profiling by LC-MS and oxalate quantification. Functional assays such as MTT, colony formation, and migration/invasion assess proliferation and metastatic potential. It is amenable to drug screening for oxalate-lowering therapies and functional genomic screens. For more information, please contact Ascent Research.
