The ALG12 Knockout A-549 Polyclonal Cells provide a heterogeneous population of CRISPR/Cas9-edited A-549 cells with disrupted ALG12 gene function. This polyclonal pool contains diverse loss-of-function alleles, eliminating clonal bias and enabling robust population-based studies of N-glycosylation. The CRISPR-mediated gene disruption targets ALG12 without introducing a predetermined mutation, generating a broad knockout model suitable for functional genomics and cellular pathway analysis.
Derived from human lung adenocarcinoma, the A-549 cell line is a canonical model for non-small cell lung cancer (NSCLC). It carries an activating KRAS G12S mutation, retains wild-type p53, and features a homozygous CDKN2A deletion, abrogating p16INK4a and p14ARF expression. This genetic profile is ideal for investigating oncogenic signaling, tumor suppression, and therapeutic resistance mechanisms in a disease-relevant epithelial context.
ALG12 encodes an alpha-1,6-mannosyltransferase that catalyzes addition of the eighth mannose residue onto the dolichol-linked oligosaccharide (DLO) precursor during N-glycan assembly in the endoplasmic reticulum. Functioning downstream of ALG9 and upstream of ALG6, ALG12 interacts with ALG3, ALG9, and the dolichol phosphate mannose synthase complex (DPM1?CDPM3). Its activity depends on dolichol phosphate and GDP-mannose availability. ALG12 disruption arrests DLO elongation, impairing N-glycosylation of nascent glycoproteins and activating ER stress pathways mediated by ATF6 and XBP1.
In A-549 cells, ALG12 knockout disrupts N-glycosylation of receptors such as EGFR, altering oncogenic signaling and epithelial cell behavior. Combined with KRAS-driven growth and CDKN2A loss, glycosylation deficiency may uncover vulnerabilities to proteotoxic stress or ER stress-inducing agents. This model thus enables dissection of how the glycocalyx and protein quality control intersect with lung cancer progression, while also serving as a cellular system for congenital disorder of glycosylation type Ig (ALG12-CDG) research.
This polyclonal knockout product supports N-glycosylation pathway dissection via LC-MS/MS glycomics, lectin blotting, and flow cytometry for cell surface glycans. Tunicamycin sensitivity and ER stress assays evaluate cellular responses, while western blotting for glycosylated proteins (ICAM-1, EGFR) validates target engagement. Transcriptional profiling of glycosylation genes by RT-qPCR measures compensatory mechanisms. Applications span cancer glycobiology, glycoproteomics, drug target validation, and CDG disease modeling. For technical inquiries, please contact Ascent Research.
