c-di-AMP Fluorinated

Difluoro 3'3'-c-di-AMP, cyclic [FdA(3’,5’)pFdA(3’,5’)p]

c-di-AMP Fluorinated (c-di[2’FdAMP]) is a synthetic analog of cyclic diadenylate monophosphate (c-di-AMP) with a fluorine atom at 2’ position of the nucleosides. c-di-AMP, a bacterial second messenger, is a potent immunostimulant in mammals. It induces production of type I interferons (IFNs) by directly binding to the endoplasmic reticulum-resident receptor STING (stimulator of interferon genes) [1, 2]. 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 [3, 4]. 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, c-di-AMP Fluorinated induces higher levels of type I IFNs than does c-di-AMP.

STING ligands such as c-di-AMP induce type I IFNs and activate interferon stimulated genes (ISG) through interferon regulatory factors (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. Jin L. et al., 2011. MPYS is required for IFN response factor 3 activation and type I IFN production in the response of cultured phagocytes to bacterial second messengers cyclic-di- AMP and cyclic-di-GMP. J Immunol. 187(5):2595-601.
2. Burdette DL. et al., 2011. STING is a direct innate immune sensor of cyclic di-GMP. Nature. 478(7370):515-8.
3. Liu P. et al., 2008. Fluorinated Nucleosides: Synthesis and biological implication. J Fluor Chem. 129(9): 743–766.
4. Böhm HJ. et al., 2004. Fluorine in medicinal chemistry. Chembiochem. 5(5):637-43.
5. Unterholzner L. et al., 2010. IFI16 is an innate immune sensor for intracellular DNA.Nat Immunol. 11(11):997-1004.
6. Zhang Z. et al., 2011. The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells. Nat Immunol.12(10):959-65.
7. Arakawa R. et al., 2010. Characterization of LRRFIP1. Biochem Cell Biol. 88(6):899-906.
8. Lippmann J. et al., 2010. IFN beta responses induced by intracellular bacteria or cytosolic DNA in different human cells do not require ZBP1 (DLM-1/DAI). Cell Microbiol. 10(12):2579-88.

Figures

THP1-Dual™ cells were stimulated for 24 hours with the STING ligands as shown (all at 10 μg/ml). IRF induction was determined by measuring the relative light units (RLUs) in a luminometer using QUANTI‑Luc™, a Lucia luciferase detection reagent.

THP1-Dual™ cells were stimulated for 24 hours with the STING ligands as shown (all at 10 μg/ml). NF‑kB induction was determined using QUANTI‑Blue™, a SEAP detection reagent, and by reading the optical density (OD) at 655 nm. Non-induced cells (NI) have been included as a negative control.

Specifications

Description: STING agonist

Synonym: 2’Fluoro-c-di-AMP sodium salt, 2',2'-di-F-c-didAMP, c-di(2’FdAMP)

CAS number: 1427269-47-3

Formula: C₂₀H₂₀F₂N₁₀O₁₀P₂ •2Na

Molecular weight: 706.36

Purity: ≥ 95% by LC/MS & NMR

Solubility: 50 mg/ml in water

Quality control:

• The ability of c-di-AMP 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

• 2x 100µg lyophilized c-di-AMP Fluorinated
• 2x 1.5 ml endotoxin-free water

Product is shipped at room temperature.

Product  should be stored at -20°C.

Product is sterile filtered prior to lyophilization.

Details