CD320 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HeLa human cervical adenocarcinoma cells, featuring targeted disruption of the CD320 gene. This heterogeneous pool of knockout cells provides a robust model for studying impaired CD320 function while minimizing clonal artifacts. Suitable for bulk biochemical assays and pooled functional screens, these cells allow investigation of transcobalamin receptor biology without single-clone bias. By avoiding single-cell cloning, the polyclonal population better represents the genetic heterogeneity inherent to tumor biology. As a genetically defined tool, they enable dissection of CD320-dependent pathways in a well-characterized cancer background.
The HeLa cell line is an immortalized epithelial model derived from cervical adenocarcinoma, widely employed in cell biology, virology, and cancer research. These cells harbor HPV-18 sequences and exhibit an aneuploid karyotype with extensive genomic rearrangements, underpinning their robust growth and experimental tractability. They have been a cornerstone of in vitro research since their isolation in 1951, contributing to breakthroughs in cancer biology, viral oncogenesis, and vaccine development. Their malignant origin and epithelial nature make them particularly relevant for studying nutrient uptake and metabolic pathway interactions in a cancer context.
CD320 encodes the transcobalamin receptor (TCblR), which mediates cellular uptake of vitamin B12 by binding and internalizing transcobalamin-cobalamin (TC-Cbl) complexes. After lysosomal degradation of the carrier, cobalamin is released and converted to methylcobalamin and adenosylcobalamin, essential cofactors for methionine synthase (MTR) and methylmalonyl-CoA mutase (MUT), respectively. These enzymes drive homocysteine remethylation and succinyl-CoA synthesis, connecting CD320 to one-carbon metabolism, DNA methylation, and nucleotide production. CD320 activity is modulated by transcobalamin availability and intracellular cobalamin levels, and it may interact with the renal receptor LRP2 in tissue-specific reabsorption.
In HeLa cancer cells, CD320 knockout provides a defined model to examine cobalamin dependency in one-carbon metabolism. Cervical adenocarcinoma cells may require robust methyl group availability for proliferation and epigenetic control; impaired CD320 is expected to reduce MTR activity, causing homocysteine and methylmalonic acid accumulation and potentially compromising nucleotide synthesis. This model allows dissection of vitamin B12 uptake effects on HeLa growth, survival, and metabolic adaptation under varied nutritional states, and facilitates exploration of CD320’s role in HPV-18-positive cancer cell chemosensitivity and epigenetic regulation.
Applications include mechanistic studies of vitamin B12 uptake, metabolic flux analysis using isotope tracers, and functional investigation of the TC-Cbl-CD320 axis. These cells support cobalamin uptake assays with radiolabeled B12, LC-MS/MS-based quantification of methylmalonic acid and homocysteine, and cell proliferation assays under B12-depleted conditions. They are also suited for drug delivery research targeting the transcobalamin pathway, in vitro modeling of methylmalonic aciduria, and genetic screens for modulators of cobalamin metabolism. Additionally, the polyclonal format enables detection of heterogeneous population responses, making it ideal for drug-response assays and resistance mechanism studies. For additional information or custom applications, contact Ascent Research.
