The GOLGA2 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the human 786-O cell line. These cells harbor targeted disruption of the GOLGA2 gene, which encodes the Golgi matrix protein GM130. This heterogeneous knockout pool provides a robust loss-of-function model for studying Golgi architecture and membrane trafficking without the constraints of clonal selection.
The 786-O cell line is a well-characterized human renal cell adenocarcinoma line originating from a clear cell renal cell carcinoma. These cells carry a mutation in the von Hippel-Lindau (VHL) tumor suppressor gene, leading to constitutive hypoxia-inducible factor activation and mimicking molecular features of ccRCC. As a kidney epithelial cell model, 786-O is widely employed for renal cancer biology, tumor metabolism, and signaling studies. The integration of GOLGA2 knockout into this VHL-mutant background offers a unique platform to examine how Golgi disruption influences oncogenic processes such as proliferation and migration.
GM130 functions as a cis-Golgi scaffold and vesicle tethering factor, essential for maintaining Golgi ribbon morphology and coordinating intercisternal trafficking. It is regulated by mitotic kinases CDK1 and PLK1, which phosphorylate GM130 to trigger Golgi disassembly during cell division. GM130 interacts directly with GRASP65 (GORASP1) and p115 (USO1) to mediate vesicle docking and stacking. Additionally, it associates with RAB1 GTPase and COPI coat components to facilitate membrane fusion and cargo sorting. Consequently, disruption of GOLGA2 leads to impaired protein secretion, defective glycosylation, and aberrant Golgi fragmentation, thereby altering vesicle-mediated transport and downstream signaling pathways.
In 786-O cells, loss of GM130 may exacerbate oncogenic phenotypes by compromising the secretory pathway and intracellular trafficking. Impaired transport of growth factor receptors, extracellular matrix components, or signaling molecules could enhance tumor cell migration and invasion. Golgi fragmentation may also affect glycosylation and processing of surface receptors, potentially sensitizing cells to targeted therapies or altering cell?Ccell communication. This model is thus invaluable for dissecting Golgi-dependent mechanisms in renal carcinoma and for identifying vulnerabilities arising from trafficking defects.
These polyclonal knockout cells are suited for a range of experimental approaches to investigate Golgi biology and cancer cell physiology. Immunofluorescence microscopy can visualize Golgi disorganization using GM130 or GRASP65 antibodies. Functional assays such as wound healing and viability tests assess migration and proliferation, while flow cytometry reveals cell cycle perturbations due to mitotic Golgi defects. Western blotting confirms GM130 loss and monitors associated proteins like p115 and RAB1. The model also supports drug screens for compounds that modulate Golgi function. For further technical details, please contact Ascent Research.
