The Sting1 Knockout RAW 264.7 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the murine macrophage RAW 264.7 line. This product features targeted disruption of the Sting1 gene, generating a validated loss-of-function model for investigating STING-dependent innate immune signaling. As a ready-to-use cell line, it enables researchers to bypass time-consuming gene-editing workflows and directly interrogate functional consequences of STING deficiency in a well-characterized host background. The cell line is supplied with quality control data demonstrating consistent growth characteristics and confirmed gene disruption, suitable for a broad range of downstream functional assays.
The host RAW 264.7 line is a BALB/c-derived macrophage cell line renowned for its robust phagocytic capacity, cytokine production, and responsiveness to Toll-like receptor and cytosolic nucleic acid agonists. These cells recapitulate key features of primary macrophages, including activation-induced morphological changes and secretion of inflammatory mediators such as tumor necrosis factor alpha (TNF-??) and interleukin-6 (IL-6). As an immortalized line, RAW 264.7 provides a genetically tractable and reproducible system for mechanistic studies of innate immunity, facilitating experiments that require homogeneous populations and scalable culture conditions. Its widespread use spans investigations of pattern recognition receptor signaling, host-pathogen interactions, and macrophage biology.
Sting1 encodes STING (stimulator of interferon genes), an endoplasmic reticulum-resident adaptor protein central to the detection of cytosolic DNA. Upon binding cyclic dinucleotides??including 2’3′-cyclic GMP-AMP (cGAMP) synthesized by cyclic GMP-AMP synthase (cGAS) in response to double-stranded DNA??STING undergoes conformational changes, traffics to the Golgi apparatus, and recruits TANK-binding kinase 1 (TBK1) and I??B kinase (IKK). This assembly triggers phosphorylation-dependent activation of interferon regulatory factor 3 (IRF3) and nuclear factor-??B (NF-??B), culminating in transcriptional induction of type I interferons (IFN-??, IFN-??) and pro-inflammatory cytokines. Additionally, STING intersects with autophagy, RIG-I, and NF-??B pathways, and interacts with regulatory proteins including TRIM56 and ULK1, underscoring its pleiotropic roles in immune responses.
In the RAW 264.7 macrophage context, disruption of STING expression selectively abolishes signaling downstream of cytosolic DNA sensors, while leaving other innate immune pathways largely intact. This model is therefore instrumental for dissecting STING-dependent versus -independent innate immune responses. It provides a defined system to evaluate the contribution of STING to macrophage activation, type I interferon production, and NF-??B-driven inflammation. The knockout cell line is particularly relevant for studying autoinflammatory conditions like STING-associated vasculopathy with onset in infancy (SAVI), systemic lupus erythematosus, and viral pathogenesis, where dysregulated STING signaling underlies disease. It also supports rational design of STING-targeted cancer immunotherapies by enabling comparative analyses with wild-type counterparts.
Typical experimental applications include stimulation with STING agonists such as cGAMP or DMXAA, followed by western blotting to assess phospho-TBK1, phospho-IRF3, and total STING levels; RT-qPCR for Ifnb1, Il6, and Tnf transcript quantification; and ELISA for secreted IFN-?? and IL-6. Additional assays encompass immunofluorescence visualization of STING translocation, flow cytometric measurement of macrophage activation markers, luciferase reporter assays for interferon-stimulated response element (ISRE) or NF-??B activity, phagocytosis assays, and multiplex cytokine profiling. This knockout model empowers rigorous dissection of STING biology in innate immunity, inflammation, and drug response. For further details or customized support, please contact Ascent Research.
