Description

The Cgas Knockout BV-2 Cell Line is a CRISPR/Cas9-edited knockout cell line that enables targeted disruption of the Cgas gene in a murine microglial background. This loss-of-function model is designed for investigators studying cytosolic DNA sensing and cGAS-dependent innate immune signaling without overexpression or dominant-negative artifacts. The knockout format provides a stable, renewable resource for repetitive experiments in neuroimmunology, virology, and neuroinflammation.

The host cell line, BV-2, is an immortalized murine microglial cell line originally derived from C57BL/6 mice. Microglia serve as the predominant immune cells of the central nervous system, continuously surveying the brain parenchyma and orchestrating neuroinflammatory responses and synaptic pruning. BV-2 cells retain key microglial features, including phagocytic capacity, responsiveness to immune stimuli, and expression of surface markers such as CD11b. This well-characterized model offers a genetically tractable system for linking cGAS function to microglial effector mechanisms.

cGAS functions as a cytosolic double-stranded DNA sensor that catalyzes the synthesis of the cyclic dinucleotide cGAMP from ATP and GTP upon binding aberrant DNA. cGAMP binds and activates the adaptor STING, which triggers TBK1-mediated phosphorylation of IRF3, leading to type I interferon production and NF-??B activation. Post-translational modifications regulate cGAS activity: acetylation by KAT5 and deacetylation by HDAC3 modulate DNA binding, while ubiquitination by RNF185 controls stability. In addition to STING and TBK1, cGAS interacts with TREX1, IFI16, and PQBP1 to fine-tune signaling. Downstream transcriptional targets include interferon-stimulated genes (e.g., ISG15, IFIT1) and autophagy-associated LC3, linking DNA sensing to innate immunity and homeostasis.

In the context of BV-2 microglia, disruption of cGAS provides a powerful tool for dissecting cell-autonomous innate immune pathways within the CNS. Since microglia are primary responders to viral infections, mitochondrial DNA release, and DNA damage-associated danger signals, the cGAS-STING axis is pivotal in triggering neuroinflammatory cascades. The knockout cell line enables researchers to determine whether microglial cGAS contributes to interferon-driven pathology in models of Aicardi-Gouti??res syndrome, systemic lupus erythematosus, or neurodegenerative disorders such as Alzheimer??s and Parkinson??s disease. Moreover, it allows assessment of cGAS involvement in microglial phagocytosis and synaptic pruning, processes that are essential for brain homeostasis and are often dysregulated in disease.

Typical applications include dsDNA transfection with cGAMP ELISA, western blotting for cGAS, STING, p-TBK1 (Ser172), and p-IRF3 (Ser396), and RT-qPCR for Ifnb1, Isg15, and Cxcl10. Immunofluorescence assays detect STING puncta and IRF3 nuclear translocation, while flow cytometry monitors CD11b and MHC-II expression. Phagocytosis assays, RNA sequencing, and cytokine bead arrays further characterize cGAS-dependent microglial functions. For further technical details, please contact Ascent Research.