The Has2 Knockout GL261 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the GL261 murine glioma cell line. This product enables disruption of the Has2 gene, which encodes hyaluronan synthase 2, the major enzyme responsible for hyaluronic acid (HA) synthesis in mammals. The polyclonal nature means it is not a single-cell-derived clone but a population of cells with heterogeneous gene edits, providing a model for studying loss-of-function effects while capturing population-level variability.
GL261 is a well-established syngeneic glioblastoma model originally induced by intracranial injection of methylcholanthrene in C57BL/6 mice. It faithfully recapitulates key features of human glioblastoma, including highly invasive growth, angiogenesis, and immune cell infiltration. Because GL261 cells are syngeneic to the C57BL/6 strain, they can be implanted orthotopically into immunocompetent mice, enabling studies of tumor?Chost immune interactions and the tumor microenvironment in a fully functional immune system.
The Has2 gene product is the principal synthase for hyaluronic acid, a major glycosaminoglycan component of the extracellular matrix. HAS2 expression is tightly controlled by upstream regulators such as EGF, TGF-??, PDGF, IL-1??, and TNF-?? via transcription factors including NF-??B, AP-1, STAT3, and HIF-1??. Once synthesized, HA binds to cell surface receptors CD44 and RHAMM, triggering downstream signaling cascades. Notably, CD44-mediated activation of PI3K?CAKT and MAPK/ERK pathways drives cell proliferation, survival, and migration. Additionally, HA signaling upregulates matrix metalloproteinases like MMP9 and promotes ??-catenin stabilization, collectively remodeling the extracellular matrix and facilitating invasive behavior.
In the context of glioblastoma, HAS2-dependent HA production is critically linked to tumor aggressiveness. Elevated HA levels in the glioma extracellular matrix foster a pro-migratory and proliferative niche, while HA?CCD44 interactions activate survival pathways and contribute to immune evasion. Disruption of Has2 in GL261 cells abolishes endogenous HA synthesis, thereby uncoupling CD44-mediated adhesion and signaling. The resulting polyclonal knockout cells offer a powerful tool to dissect how HA depletion alters glioma cell behavior, matrix dynamics, and response to therapeutic interventions.
Research applications for this knockout model are broad. It is suited for examining the role of hyaluronic acid in the glioblastoma microenvironment, evaluating CD44 and RHAMM-dependent signaling cascades, and testing inhibitors of hyaluronan synthesis. The cells are compatible with standard assays such as hyaluronic acid ELISA, western blotting for HAS2 protein, RT?qPCR for Has2 transcript, transwell migration and invasion assays, proliferation assays, flow cytometry for CD44 surface expression, and immunofluorescence staining of the extracellular HA matrix. In addition, syngeneic orthotopic implantation in C57BL/6 mice enables in vivo assessment of tumor growth, invasion, and immune cell infiltration. For further technical details, please contact Ascent Research.
