The GOLGA2 Knockout HEK293T Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of human embryonic kidney HEK293T cells carrying targeted disruption of the GOLGA2 gene. This polyclonal knockout pool provides a genetically heterogeneous loss-of-function model, enabling robust analysis of GOLGA2-dependent processes without single-cell cloning artifacts. The gene-edited population is suitable for studying Golgi apparatus organization, vesicle-mediated transport, and cell polarity regulation.
HEK293T cells are an extensively used mammalian host cell line derived from human embryonic kidney epithelial cells, immortalized by stable expression of the SV40 large T antigen. These cells support high-level transient protein expression and efficient lentiviral vector production, making them a versatile platform for biochemical, cell biological, and virological investigations. Their epithelial origin and robust growth characteristics also facilitate studies of cell architecture, secretion, and migration.
GOLGA2 encodes GM130, a cis-Golgi matrix protein essential for maintaining Golgi ribbon integrity and mediating vesicle tethering. GM130 interacts with USO1/p115 and GORASP1/GRASP65 to organize the Golgi stack and functions downstream of RAB1 GTPase and ARF1 during ER-to-Golgi transport. Its activity is regulated by CDK1-mediated phosphorylation during mitosis, which drives Golgi disassembly. Knockout of GOLGA2 disrupts downstream targets including Golgi ribbon structure, protein secretion and glycosylation, cell surface protein localization, and directed cell migration, thereby affecting Golgi organization, ER-to-Golgi vesicle-mediated transport, mitotic Golgi fragmentation, and cell polarity signaling pathways.
In HEK293T cells, loss of GM130 disrupts Golgi morphology and compromises the fidelity of protein trafficking and post-translational modification, providing a powerful model to dissect Golgi-dependent processes. This knockout model enables precise investigation of how Golgi fragmentation influences protein secretion efficiency, glycoprotein processing, and cell surface receptor presentation. Additionally, it allows exploration of the interplay between Golgi structure and epithelial cell polarity, as well as the role of GM130 in directed cell migration??a process frequently dysregulated in cancer metastasis.
Researchers can employ this polyclonal knockout pool in a wide array of experimental applications, including immunofluorescence microscopy to visualize Golgi fragmentation, Western blotting to confirm GM130 depletion and assess Golgi markers, and SEAP or other secretion assays to quantify protein trafficking efficiency. Lectin-based glycosylation analysis can reveal alterations in glycan processing, while cell migration and invasion assays probe the functional consequences of GM130 loss on motility. Co-immunoprecipitation studies with known interacting partners such as USO1/p115 or GORASP1/GRASP65 can further elucidate molecular interactions. These applications support fundamental studies in Golgi biology, trafficking, and disease modeling, including Golgi fragmentation-related neurodegenerative disorders and cancer cell migration. For additional information or customized solutions, please contact Ascent Research.
