The GOLIM4 Knockout A-549 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted disruption of the GOLIM4 gene in the human A-549 lung adenocarcinoma epithelial cell line. This polyclonal format provides a heterogeneous pool of edited cells, enabling robust loss-of-function studies without clonal selection bias. The product serves as a versatile tool for investigating Golgi membrane biology, glycosylation pathways, and toxin?Chost interactions in a disease-relevant cellular context.
A-549 cells are derived from lung adenocarcinoma tissue of a 58-year-old male and harbor a KRAS G12S driver mutation. These cells are widely established as a model of alveolar type II epithelium, exhibiting characteristics of surfactant production and epithelial barrier function. Their malignant origin and well-characterized signaling landscape make them particularly suitable for studying oncogenic glycosylation changes, metal homeostasis, and endocytic trafficking in a cancer cell background.
GOLIM4 encodes a pH-sensitive type II Golgi membrane protein that cycles between the Golgi apparatus and endosomes, where it functions as a cargo receptor and contributes to retrograde transport, protein glycosylation, and manganese detoxification. It is regulated by Golgi luminal pH and the small GTPase ARF1, and it physically interacts with the conserved oligomeric Golgi (COG) complex and the Shiga toxin B subunit. GOLIM4 operates downstream of ARF1 and upstream of cell surface glycoprotein maturation and lysosomal protein sorting. Within the retrograde trafficking pathway, it cooperates with RAB6, VPS52, and the manganese transporter TMEM165. Disruption of GOLIM4 ablates the cell surface receptor for Shiga toxin, thereby conferring cellular resistance to the toxin, while simultaneously impairing manganese efflux and increasing sensitivity to manganese-induced cytotoxicity.
In the A-549 KRAS-mutant adenocarcinoma setting, GOLIM4 knockout provides a clinically relevant model for examining the interplay between oncogenic signaling and Golgi-dependent pathways. Loss of GOLIM4-mediated trafficking is expected to perturb glycocalyx composition and the presentation of tumor-associated carbohydrate antigens, offering insights into immune evasion and metastasis. The dual phenotype of Shiga toxin resistance and manganese hypersensitivity uniquely positions this model for pharmacological and toxicological investigations, including studies on Shiga toxin-associated hemolytic uremic syndrome and manganese neurotoxicity.
Typical research applications encompass functional characterization of Golgi trafficking, glycobiology, and retrograde transport mechanisms. Researchers can employ Shiga toxin sensitivity assays and manganese toxicity assays to quantify phenotypic responses, western blotting and immunofluorescence to assess glycosylation states and Golgi morphology, and glycoproteomics to profile cell surface glycan alterations. RT-qPCR can be used to monitor compensatory changes in COG complex subunits or TMEM165 expression. This polyclonal knockout population is also suitable for pooled CRISPR screens and high-content imaging-based trafficking assays. For further information, please contact Ascent Research.
