The ALG3 Knockout A-549 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population originating from the human A-549 lung adenocarcinoma line. This product delivers a loss-of-function model for ALG3, the gene encoding dolichyl-PP-Man:Man5GlcNAc2-PP-dolichol alpha-1,3-mannosyltransferase, a key enzyme in N-linked glycan precursor assembly. The polyclonal nature allows direct analysis of gene disruption effects without clonal expansion, facilitating efficient functional studies.
A-549 cells were established from a lung adenocarcinoma of a 58-year-old Caucasian male and serve as a widely used model for human lung adenocarcinoma and respiratory epithelial barrier studies. This adherent epithelial line retains alveolar type II-like traits, providing a consistent and well-characterized host for knockout experiments. Its robust growth and defined signaling pathways make it suitable for exploring glycosylation-related processes.
ALG3 catalyzes the transfer of the first mannose residue to Man5GlcNAc2-PP-dolichol in the endoplasmic reticulum (ER), a critical commitment step in dolichol-linked oligosaccharide biosynthesis. This reaction precedes elongation by ALG9, ALG12, and ALG6 within the oligosaccharyltransferase assembly complex. ALG3 disruption leads to accumulation of truncated lipid-linked oligosaccharides and global hypoglycosylation of N-glycosylated secretory and membrane proteins, thereby activating the unfolded protein response (UPR) and ER stress pathways. The ALG3-dependent pathway is interconnected with ER protein processing and quality control, influencing downstream cellular homeostasis.
In the A-549 adenocarcinoma context, ALG3 knockout illuminates links between aberrant N-glycosylation and lung cancer phenotype. Hypoglycosylated adhesion molecules and receptors may alter cell signaling, immune evasion, and metastatic behavior. Concomitant ER stress provides a model to study glycosylation-dependent survival mechanisms and UPR activation in cancer cells. Additionally, as ALG3 mutations underlie congenital disorder of glycosylation type Id (ALG3-CDG), these cells offer a surrogate system to investigate molecular pathologies of CDG, complementing studies in neurodevelopmental models.
Typical applications include western blotting and lectin blotting for glycoprotein profiling, RT-qPCR for UPR markers (e.g., HSPA5, DDIT3), flow cytometry with lectins for surface glycan analysis, immunofluorescence for ER stress markers, metabolic labeling of glycoproteins, and N-glycan mass spectrometry. The model supports research into N-glycan biosynthesis, ER stress, cancer glycobiology, and congenital disorders of glycosylation. These cells are also suitable for investigating how ALG3 loss affects drug sensitivity or epithelial barrier function. For further information or customized solutions, please contact Ascent Research.
