CD320 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population with targeted disruption of the CD320 gene in the human SK-HEP-1 cell line. This loss?of?function model abolishes transcobalamin receptor (TCblR) expression, enabling dissection of cobalamin (vitamin B12) uptake and downstream metabolism without clonal selection artifacts.
SK-HEP-1 is an ascites?derived human hepatic adenocarcinoma line with adherent, endothelial?like characteristics. It is a widely used surrogate for liver sinusoidal endothelial cells and a standard host for hepatic drug metabolism studies, combining hepatocyte-like and vascular features.
CD320 encodes TCblR, which specifically internalizes the transcobalamin II (TCN2)?Ccobalamin complex. Internalized cobalamin acts as a cofactor for cytosolic methionine synthase (MTR) and mitochondrial methylmalonyl?CoA mutase (MMUT). MTR remethylates homocysteine to methionine within folate?mediated one?carbon metabolism, while MMUT converts methylmalonyl?CoA to succinyl?CoA in propionate metabolism. CD320 function is influenced by TCN2, cobalamin levels, and the LRP2 endocytic receptor.
In the hepatic context, CD320 knockout recapitulates features of transcobalamin receptor deficiency and methylmalonic aciduria, marked by elevated methylmalonic acid and homocysteine. The SK-HEP-1 background provides a relevant setting to study liver-specific contributions to systemic B12 handling, one-carbon unit distribution, and sinusoidal uptake mechanisms.
These polyclonal knockout cells support functional assays including cobalamin uptake kinetics, MTR activity measurements, methylmalonic acid quantification, and homocysteine profiling. Researchers can validate CD320 disruption via western blot or RT?qPCR, perform immunofluorescence localization, and assess viability under B12 deprivation. They are ideal for isogenic rescue and transcriptomic comparisons. For further details, contact Ascent Research.
