CD320 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the A-549 lung adenocarcinoma cell line, with disruption of the CD320 gene. CD320 encodes the transcobalamin receptor that mediates cellular uptake of vitamin B12 (cobalamin) via transcobalamin II (TCN2). This knockout product provides a heterogeneous loss-of-function model for studying the effects of CD320 ablation on cobalamin internalization and downstream metabolic pathways. As a polyclonal population, it is suitable for bulk-functional assays without clonal selection.
A-549 cells are a widely used human lung adenocarcinoma model, established from an adult male patient. They exhibit alveolar type II pneumocyte characteristics, including surfactant production and epithelial morphology, and serve as a standard system for non-small cell lung cancer research. Their robust growth and ease of genetic manipulation make them an ideal host for CRISPR-based gene editing.
CD320 functions as a receptor for TCN2-bound cobalamin, mediating its clathrin-dependent endocytosis in concert with LRP2 (megalin) and adaptor protein AP2. Internalized cobalamin acts as a cofactor for methionine synthase (MTR) and methylmalonyl-CoA mutase (MUT), key enzymes in one-carbon metabolism and methylmalonate catabolism. CD320 expression is transcriptionally regulated by SP1, and its activity influences homocysteine remethylation, methylmalonic acid clearance, and S-adenosylmethionine-dependent DNA methylation. Disruption of CD320 impairs B12 uptake, leading to accumulation of homocysteine and methylmalonic acid and altered methionine synthesis.
In A-549 cells, CD320 knockout provides a model to explore the intersection of vitamin B12 metabolism and lung adenocarcinoma biology. These cancer cells often rely on one-carbon metabolism for proliferation and nucleotide synthesis, making them vulnerable to disruptions in cobalamin handling. This system permits investigation of how impaired B12 uptake affects tumor growth, DNA methylation patterns, and metabolic flexibility, and can reveal compensatory pathways or therapeutic targets.
Applications include cobalamin uptake assays, homocysteine and methylmalonic acid quantification, immunoblotting for MTR and MUT, and RT-qPCR for CD320 verification. Further studies involve proliferation under B12 limitation, DNA methylation analysis, and metabolic flux profiling. These cells are suited for research on cancer metabolism, epigenetic regulation, and lung adenocarcinoma pathogenesis. For technical inquiries, please contact Ascent Research.
