C1GALT1C1 Knockout A-549 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma epithelial cell line. This product features targeted disruption of the C1GALT1C1 gene, encoding the COSMC molecular chaperone, introduced by CRISPR/Cas9 genome editing to create a loss-of-function model for investigating core 1 O-glycosylation. The polyclonal nature provides a heterogeneous population with a spectrum of gene-editing outcomes, enabling researchers to study biological processes influenced by C1GALT1C1 expression levels without clonal selection bias.
The A-549 host cell line was originally isolated from a lung adenocarcinoma tumor of a 58-year-old male donor and serves as a widely employed epithelial model in cancer biology and glycobiology. These adherent cells exhibit characteristics of alveolar type II epithelium and retain key features of malignant transformation, including altered glycosylation patterns. A-549 cells are known to express mucins and other glycoproteins, making them an ideal platform to examine the role of O-glycan elongation in epithelial tumor cell behavior.
C1GALT1C1 (COSMC) operates as a dedicated endoplasmic reticulum chaperone essential for the correct folding and functional maturation of C1GALT1 (core 1 beta3-galactosyltransferase/T-synthase), the enzyme responsible for generating core 1 O-glycans (T antigen). In the absence of functional COSMC, C1GALT1 misfolds and undergoes degradation, leading to loss of T-synthase activity and accumulation of the truncated precursor Tn antigen on glycoproteins. Upstream regulators, including the unfolded protein response sensors ATF6, IRE1, and PERK, can influence COSMC expression under ER stress. Downstream, COSMC deficiency destabilizes C1GALT1 protein and disrupts O-glycosylation of key substrates such as MUC1 and podoplanin. Thus, COSMC acts as a gatekeeper in the O-glycan biosynthesis pathway, with its disruption redirecting glycosylation toward Tn and sialyl-Tn antigen formation.
In the A-549 lung carcinoma background, ablation of C1GALT1C1 profoundly alters the cell-surface glycome, mimicking the aberrant glycosylation commonly observed in human cancers. This model enables the investigation of how truncated O-glycans influence various oncogenic processes, including altered cell adhesion, migration, and immune recognition, which are often linked to Tn antigen expression. Furthermore, the epithelial origin of these cells allows the study of mucin-type glycoprotein function in the context of lung adenocarcinoma, providing insights into glycocalyx-mediated signaling and metastasis. The polyclonal knockout format simulates the heterogeneous glycosylation profiles seen in clinical tumors, offering a more clinically relevant experimental system than a clonally selected knockout line.
These C1GALT1C1 knockout polyclonal cells enable diverse glycobiology applications, including Tn antigen detection via Vicia villosa lectin immunofluorescence or flow cytometry, western blotting for C1GALT1 and under-glycosylated targets, RT-qPCR verification, O-glycan mass spectrometry profiling, cell adhesion assays, and lectin blotting. The model is invaluable for investigating COSMC function in cancer-associated aberrant glycosylation, mucin biology, and Tn antigen-driven phenotypic changes. For further information or to order, please contact Ascent Research.
