2'3'-cGAMP Control

STING agonist negative control

2’3’-cGAMP Control, also known as 2’5’-GpAp, is a linear dinucleotide analog after hydrolysis of cyclic guanosine monophosphate- adenosine monophosphate (2’3’-cGAMP) by phosphodiesterases [1].

2’3’-cGAMP is a cyclic dinucleotide (CDN) produced in mammalian cells by cGAS (cGAMP synthase) in response to cytosolic DNA. CDNs including 2’3’-cGAMP bind the cytosolic DNA sensor STING (stimulator of interferon genes) and induce the production of type I interferons (IFNs).

Due to its linear conformation, 2’5’GpAp is intended to serve as a negative control for 2’3’-cGAMP in type I IFN induction assays.

The importance of dinucleotide conformation has been well established in bacteria, where RNA regulatory riboswitches are able to discriminate between biologically active CDNs and their corresponding linear dinucleotides [1-3].

To facilitate the study of CDNs and the IFN pathway, InvivoGen has developed stable reporter cells in the human monocytic THP-1 cell line. These cells express a reporter gene (either SEAP or the secreted Lucia luciferase) under the control of an interferon regulatory factors (IRF)-inducible promoter.

As expected, 2’3’-cGAMP Control does not induce a type I IFN response in this immune cell model.

References:

1. Gao J. et al., 2015. Identification and characterization of phosphodiesterases that specifically degrade 3'3'-cyclic GMP-AMP. Cell Res. 25(5):539-50
2. Ren A. et al., 2015. Structural Basis for Molecular Discrimination by a 3',3'-cGAMP Sensing Riboswitch. Cell Rep. 11(1):1-12.
3. Smith K. et al., 2012. Structural and biochemical characterization of linear dinucleotide analogues bound to the c-di-GMP-I aptamer. Biochemistry. 51(1):425-32.
4. Unterholzner L. et al., 2010. IFI16 is an innate immune sensor for intracellular DNA.Nat Immunol. 11(11):997-1004.
5. 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.
6. Arakawa R. et al., 2010. Characterization of LRRFIP1. Biochem Cell Biol. 88(6):899-906.
7. Lippmann J. et al., 2010. IFNbeta 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.

Specifications

Source: Synthetic

Activity: 2'3'-cGAMP negative control

Synonym: 2'5'-GpAp

Formula: C₂₀H₂₃N₁₀O₁₄P₂.3Na

Molecular weight: 758.38

Solubility: 50 mg/ml in water

Quality control:

• Purity and structure have been determined by LC/MS and NMR: ≥ 95%
• The inability of 2'3'-cGAMP Control to induce type I interferon (IFN) has been confirmed in cellular assays.
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.

Contents

• 1 mg of lyophilized chemically synthesized 2’3’-cGAMP Control (2’5’-GpAp)
  Note: 2’3’-cGAMP Control is sterile filtered prior to lyophilization.
• 1.5 ml endotoxin-free water.

Product is shipped at room temperature.

Stored at -20 °C.

Details