3'3'-cGAMP Fluorinated

STING Ligand - Difluoro 3'3'-cGAMP, cyclic [FdG(3’,5’)pFdA(3’,5’)p]

3’3’-cGAMP Fluorinated (c-[2'FdGMP]-[2'FdAMP]) is a synthetic analog of cyclic guanosine monophosphate- adenosine monophosphate (cyclic GMP-AMP, cGAMP) with a fluorine atom at 2’ position of the nucleosides. 3’3’-cGAMP is a cyclic di-nucleotide produced by bacteria. It is also referred to as "canonical" cGAMP due the presence of the classical 3’-5’ phosphodiester linkages between the guanosine and the adenosine. It has been reported that cGAMP binds STING (stimulator of IFN genes) and subsequently induces TBK1-IRF3-dependent production of IFN-β [1].

The incorporation of fluorine into biologically active molecules is commonly used in medicinal chemistry to improve their metabolic stability or to modulate physicochemical properties such as lipophilicity [2, 3]. Moreover, the introduction of a fluorine atom can change the biological activities of a molecule. Interestingly, when used at low concentrations in various cellular assays, 3’3’-cGAMP Fluorinated induces higher levels of type I IFNs than does cGAMP.

STING ligands such as cGAMP induce type I IFNs and activate interferon stimulated genes (ISG) through IRFs. To facilitate their study, InvivoGen has developed stable reporter cells in two well established immune cell models: THP-1 human monocytes and RAW 264.7 murine macrophages. These cells express a reporter gene (SEAP or Lucia luciferase), under control of an IRF-inducible promoter.

References:

1. Zhang X. et al., 2013. Cyclic GMP-AMP containing mixed phosphdiester linkages is an endogenous high-affinity ligand for STING. Mol Cell.51(2):226-35.
2. Liu P. et al., 2008. Fluorinated Nucleosides: Synthesis and biological implication. J Fluor Chem. 129(9): 743–766.
3. Böhm HJ. et al., 2004. Fluorine in medicinal chemistry. Chembiochem. 5(5):637-43.

Figures

THP1-Dual™ cells were stimulated for 24 hours with the STING ligands as shown (all at 10 μg/ml). The interferon regulatory factor (IRF) response was assessed by measuring the activity of Lucia luciferase in the supernatant using QUANTI-Luc™ 4 Lucia/Gaussia. Data are shown in fold response over non-induced cells (mean ± SEM).

THP1-Dual™ cells were stimulated for 24 hours with the STING ligands as shown (all at 10 μg/ml). The NF-κB-induced SEAP activity was assessed using QUANTI‑Blue™ Solution, a SEAP detection reagent. Data are shown as optical density (OD) at 630 nm (mean ± SEM). Non‑induced cells (NI) have been included as a negative control.

Specifications

Description: STING agonist

Synonym: 2’Fluoro-cyclic GMP-AMP, c-(2'FdGMP)-(2'FdAMP), F-c-GpAp sodium salt

Formula: C₂₀H₂₀F₂N₁₀O₁₁P₂ •2Na

Molecular weight: 722.36

Purity: ≥ 95% by LC/MS and NMR

Solubility: 50 mg/ml in water

Quality control:

• The ability of 3’3’-cGAMP Fluorinated to induce type I interferon (IFN) has been confirmed using cellular assays.
• The absence of endotoxins and other bacterial contaminants has been confirmed using HEK-Blue™ TLR4 and HEK-Blue™ TLR2 cells.

Contents

• 5 x 100 µg lyophilized 3’3’-cGAMP Fluorinated
• 2 x 1.5 ml endotoxin-free water

Product is shipped at room temperature.

Upon receipt, store at -20°C.

Details