The Vsir Knockout BV-2 Cell Line is a CRISPR/Cas9-edited knockout cell line generated from the immortalized mouse microglial BV-2 cell line. This product provides a powerful loss-of-function model for investigating the role of Vsir, which encodes the V-domain Ig suppressor of T cell activation (VISTA), in central nervous system (CNS) immune regulation. The cell line is designed for researchers studying immune checkpoint signaling, neuroinflammation, and microglial biology. Using CRISPR/Cas9-mediated gene disruption, stable ablation of Vsir has been achieved, enabling detailed functional dissection of VISTA-dependent pathways in a microglial context.
The BV-2 host cell line is an immortalized mouse microglial cell line originally derived from C57BL/6 mice. Microglia are the resident immune cells of the CNS, responsible for immune surveillance, phagocytosis, and the regulation of neuroinflammatory responses. BV-2 cells retain key features of primary microglia, including responsiveness to inflammatory stimuli such as lipopolysaccharide (LPS) and interferon-gamma (IFN-??), and are widely used as a model system for microglial biology. This cell line expresses microglial markers and exhibits phagocytic activity, making it a relevant platform for studying neuroinflammatory processes in vitro.
Vsir encodes VISTA, an immune checkpoint molecule that functions as a negative regulator of T-cell activation and cytokine production. VISTA binds to its ligand PSGL-1 to deliver inhibitory signals that suppress T-cell receptor-mediated activation, reduce pro-inflammatory cytokine release, and promote regulatory T-cell differentiation. In the BV-2 microglial system, VISTA is poised to modulate key signaling cascades, including the NF-??B pathway and the PI3K/AKT/mTOR axis. Mechanistically, VISTA signaling involves recruitment of SHP-1 and SHP-2 phosphatases, which dephosphorylate downstream effectors, leading to attenuated NF-??B and PI3K/AKT activity, thereby reducing expression of inflammatory mediators such as TNF-??, IL-6, and IL-2. Upstream regulators like IFN-??, IL-10, and TLR ligands can modulate Vsir expression, integrating VISTA into broader immune-regulatory networks.
In the context of microglial cells, knockout of Vsir allows researchers to dissect how VISTA governs neuroinflammatory responses. Microglial activation is a hallmark of many neurodegenerative and neuroinflammatory diseases, including Alzheimer??s disease, Parkinson??s disease, and multiple sclerosis. By disrupting Vsir, this cell line enables the study of VISTA??s role in regulating microglial phagocytosis, cytokine secretion, and migration. The model is particularly valuable for investigating the crosstalk between immune checkpoint pathways and neuroinflammation, providing insights into potential therapeutic targets for CNS disorders where microglial dysregulation plays a critical role.
Typical research applications include neuroinflammation studies, immune checkpoint regulation in the CNS, and microglial activation assays. Researchers can employ qRT-PCR to confirm Vsir knockout, flow cytometry to assess microglial activation markers such as CD68 and Iba1, and ELISA to quantify secreted cytokines like TNF-?? and IL-6. Western blotting can be used to monitor VISTA protein levels and downstream signaling events, including phosphorylation of AKT and NF-??B. Phagocytosis assays and live-cell imaging for migration further extend the utility of this cell line. For additional technical details or to discuss custom applications, please contact Ascent Research.
