The Cgas Knockout B16-F10 Cell Line is a CRISPR/Cas9-edited mouse melanoma cell line featuring targeted disruption of the Cgas gene, which encodes cyclic GMP-AMP synthase (cGAS). This product provides a stable loss-of-function model in the highly metastatic B16-F10 background, enabling precise investigation of cGAS-dependent innate immune signaling in cancer biology. The knockout cell line is suitable for a wide range of in vitro and in vivo experimental systems, including syngeneic tumor studies in C57BL/6 mice, and is maintained under standard culture conditions without antibiotic selection pressure.
The parental B16-F10 cell line is a well-characterized subline of the B16 melanoma originally isolated from a spontaneous tumor in C57BL/6J mice. It was specifically selected for enhanced lung colonization potential, making it a robust model for studying melanoma metastasis and tumor?Chost interactions. B16-F10 cells exhibit aggressive growth kinetics and are syngeneic to the C57BL/6 background, facilitating immunocompetent transplantation experiments that preserve the tumor microenvironment and immune contexture. This host line is routinely employed to evaluate immunotherapeutic strategies, metastatic dissemination, and mechanisms of immune evasion.
cGAS functions as a primary cytosolic double-stranded DNA sensor that, upon binding DNA from sources such as micronuclei, mitochondrial leakage, or viral genomes, catalyzes the synthesis of the second messenger cyclic GMP-AMP (cGAMP). cGAMP subsequently binds and activates the endoplasmic reticulum-resident adaptor protein STING (TMEM173), triggering its trafficking to the Golgi apparatus. There, STING recruits and activates TANK-binding kinase 1 (TBK1) and IKK kinases, leading to phosphorylation of interferon regulatory factor 3 (IRF3) and activation of nuclear factor-??B (NF-??B), respectively. These events culminate in transcriptional induction of type I interferons (IFN-??, IFN-??) and pro-inflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-?? (TNF-??), along with interferon-stimulated genes including ISG15 and CXCL10. The pathway is tightly regulated by factors such as the exonuclease TREX1, which degrades cytosolic DNA, and by interacting partners like PQBP1, ZCCHC3, and IFI16, which modulate cGAS activity or DNA recognition.
In the context of B16-F10 melanoma, the cGAS-STING axis is a critical determinant of tumor immunogenicity and metastatic progression. cGAS-mediated detection of tumor-derived DNA can initiate innate immune responses that shape the tumor microenvironment, influencing immune cell infiltration and anti-tumor immunity. Disruption of Cgas in this highly metastatic line permits dissection of how loss of DNA sensing alters cytokine profiles, STING-dependent signaling, and susceptibility to immune-mediated clearance. Moreover, the knockout model is invaluable for dissecting the contributions of host-derived cGAS versus tumor-intrinsic cGAS in syngeneic grafting experiments, thereby clarifying mechanisms of immune surveillance and escape in melanoma.
This knockout cell line supports diverse research applications, including the study of innate immune sensing in melanoma, the role of cGAS-STING in regulating the immunosuppressive tumor microenvironment, and the evaluation of therapeutic strategies such as STING agonists and DNA-damaging agents like chemotherapy or radiation. Researchers can employ a variety of representative assays, including western blotting for cGAS, STING, phospho-TBK1, and phospho-IRF3; RT-qPCR for Ifnb1, Isg15, and Cxcl10; ELISA for cGAMP and cytokine quantification; immunofluorescence to visualize STING trafficking; and flow cytometry to assess immune cell infiltration. Syngeneic implantation in C57BL/6 mice combined with tumor growth and metastasis analyses further extends the model??s utility. For additional details, please contact Ascent Research.
