The CD320 Knockout A2780 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal cell population derived from the A2780 human ovarian carcinoma line, in which the CD320 gene has been disrupted to abrogate its function. This loss-of-function model is designed for investigations into vitamin B12 trafficking and the downstream metabolic and epigenetic consequences in a cancer-relevant background. The polyclonal format preserves a heterogeneous knockout pool to study population-level effects of CD320 disruption without clonal selection biases, suitable for pooled functional genomics or pharmacological screening applications.
The parental A2780 cell line was established from an untreated patient with ovarian endometrioid adenocarcinoma and exhibits an adherent epithelial morphology. Widely employed as a model of ovarian cancer, A2780 cells retain key characteristics of the disease and are recognized for their utility in drug resistance, signal transduction, and metabolic studies. This genetic background provides a clinically relevant context for dissecting how cobalamin utilization influences tumor cell biology, as ovarian carcinomas may alter nutrient uptake pathways to sustain proliferation.
CD320 encodes the receptor that binds the transcobalamin II (TCN2)-cobalamin complex and mediates its endocytosis, a critical step for intracellular cobalamin delivery. It interacts with clathrin adaptor proteins such as LDLRAP1 for uptake and is regulated by SREBP transcription factors, vitamin B12 status, and TNF-??. Cobalamin acts as a cofactor for methionine synthase (MTR) and methylmalonyl-CoA mutase (MUT), driving methionine and succinyl-CoA synthesis and regulating homocysteine and methylmalonic acid levels. These enzymes intersect with one-carbon metabolism through MTHFR and MTRR, positioning CD320 as a key regulator of methylation and mitochondrial function.
In A2780 ovarian cancer cells, CD320 knockout disrupts intracellular cobalamin homeostasis, limiting MTR and MUT activities and elevating homocysteine and methylmalonic acid. This metabolic shift can impair one-carbon metabolism, nucleotide synthesis, and epigenetic methylation, potentially affecting tumor proliferation, stemness, or drug response. The model thus enables dissection of whether cobalamin-dependent pathways sustain ovarian carcinoma growth, and how accumulation of toxic intermediates may reveal cancer-specific vulnerabilities. It also provides a human cell-based system for studying CD320-linked disorders like methylmalonic aciduria.
Research applications include investigating vitamin B12 trafficking defects in cancer, the impact of cobalamin metabolism on ovarian cancer proliferation and epigenetic regulation, and modeling CD320-deficiency syndromes. Typical assays range from western blotting and RT-qPCR for target verification to LC-MS quantification of homocysteine and methylmalonic acid and enzymatic activity measurements for MTR and MUT. Functional readouts such as MTT-based proliferation, Annexin V apoptosis staining, RNA-seq transcriptomics, and drug sensitivity screening with B12 analogs further enable comprehensive pathway analysis. For additional information or technical support, please contact Ascent Research.
