The CD320 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma cell line. This product features targeted disruption of the CD320 gene, generating a loss-of-function model for the transcobalamin II (TCN2)-cobalamin receptor. The polyclonal nature ensures representation of diverse gene disruption events, providing a robust system for studying CD320-dependent processes without reliance on single-cell clones.
The Huh-7 host cell line was established from a well-differentiated hepatocellular carcinoma of a 57-year-old male and retains hepatocyte-like functions, including secretion of plasma proteins and bile acids. Widely employed for hepatitis C virus replication studies and liver cancer research, Huh-7 offers a well-characterized metabolic background for interrogating nutrient transporter biology.
CD320 encodes the receptor for the TCN2-cobalamin complex, mediating its clathrin-dependent endocytosis. The delivered vitamin B12 acts as cofactor for methionine synthase (MTR) and methylmalonyl-CoA mutase (MUT), driving homocysteine remethylation and methylmalonic acid catabolism. Key interacting factors include TCN2, clathrin, and adaptor protein complex 2 (AP-2), positioning CD320 upstream of one-carbon metabolism and mitochondrial function.
In Huh-7 hepatocellular carcinoma cells, CD320 knockout allows exploration of impaired vitamin B12 uptake on liver cancer biology. Hepatocytes are primary sites of cobalamin storage and utilization; CD320 loss mimics deficiency states, disrupting methylation, nucleotide synthesis, and redox balance??processes frequently altered in hepatocarcinogenesis. This model helps elucidate CD320-dependent metabolic roles in tumor progression.
Applications include investigating vitamin B12 uptake deficiency, studying effects of impaired one-carbon metabolism in hepatocellular carcinoma, modeling methylmalonic aciduria in liver context, and assessing cobalamin’s role in methylation and nucleotide synthesis. Representative assays comprise cobalamin uptake measurements, homocysteine and methylmalonic acid quantification, methionine synthase activity, cell viability under B12 restriction, and metabolomics. For further information or to obtain these cells, please contact Ascent Research.
