The CD320 Knockout 786-O Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt CD320 gene expression in the 786-O human renal cell adenocarcinoma line. This loss-of-function model provides researchers with a powerful tool to investigate the transcobalamin receptor in a well-established clear cell renal cell carcinoma (ccRCC) background. The polyclonal format captures a heterogeneous array of gene-disrupted alleles without single-cell cloning, offering a realistic representation of knockout effects across a cell population and avoiding clonal bias. Standard validation approaches include Western blotting and RT-qPCR to confirm CD320 knockout, and flow cytometry to assess surface receptor loss, enabling robust phenotypic comparisons with unedited controls.
The 786-O host cell line originates from human renal cell adenocarcinoma and is widely used as a ccRCC model. It harbors a homozygous inactivating mutation in the VHL tumor suppressor gene, leading to constitutive stabilization of hypoxia-inducible factor 2-alpha (HIF-2??) under normoxic conditions. This genetic backdrop mimics pseudohypoxic signaling, driving a metabolic and angiogenic phenotype characteristic of aggressive ccRCC. Consequently, 786-O cells provide a physiologically relevant platform for studying oncometabolism, tumor progression, and therapeutic vulnerabilities, particularly in the context of altered micronutrient dependence.
CD320 encodes the cell surface receptor that binds transcobalamin II (TCN2)-cobalamin complexes, enabling receptor-mediated endocytosis of vitamin B12. Upon internalization, cobalamin serves as a cofactor for two key enzymes: methionine synthase (MTR), which converts homocysteine to methionine in the methionine cycle, and methylmalonyl-CoA mutase (MUT), which processes methylmalonyl-CoA to succinyl-CoA in the mitochondria. These reactions couple one-carbon metabolism with nucleotide synthesis, methylation reactions, and energy production. CD320 function is regulated upstream by cellular cobalamin status and hypoxia-inducible factors, while downstream targets include MTR, MUT, and the broader methylation network. Disruption of CD320 thus decouples B12 sensing from these essential metabolic pathways, allowing detailed dissection of cobalamin-dependent signaling.
In the 786-O ccRCC context, CD320-mediated cobalamin uptake may intersect with HIF-2??-driven metabolic reprogramming. Renal tumor cells often exhibit heightened one-carbon metabolism to support proliferation and redox balance, creating a potential reliance on vitamin B12 transport. Knocking out CD320 enables investigation of how cobalamin availability influences tumor proliferation, migration, invasion, and sensitivity to therapeutics such as methotrexate or nucleoside analogs. Since the polyclonal population reflects a spectrum of gene-disruption events, it is particularly suited for functional assays that interrogate the collective impact of CD320 loss without the artifacts of single-clone selection, enhancing translational relevance.
This product is ideally suited for a range of biomedical research applications, including functional analysis of cobalamin metabolism in renal carcinoma, characterization of CD320-driven tumor phenotypes, and examination of vitamin B12 dependency in ccRCC. Representative assays include proliferation and migration/invasion studies, B12 uptake quantification, homocysteine and methylmalonic acid measurement to gauge enzyme activity, methionine synthase activity assays, and drug sensitivity profiling. These experiments can be coupled with Western blotting, RT-qPCR, and flow cytometry for molecular validation. For further technical details or ordering information, please contact Ascent Research.
