c-di-AMP disodium

• CAS: 2734909-87-4
• Catalog Number: ACM-ONT2734909874
• Category: Cyclic Dinucleotides
• Molecular Weight: 702.38
• Molecular Formula: C20H22N10Na2O12P2

Case Study

Cyclic di-AMP for Investigating Thymineless Death Regulation in Listeria monocytogenes

Leeming JP, et al. mBio, 2025, 17(1).

Cyclic di-AMP was employed as a key molecular regulator to investigate thymineless death (TLD) in Listeria monocytogenes. In this study, intracellular c-di-AMP levels were genetically manipulated in a ΔthyA mutant to evaluate bacterial survival under thymidine starvation. Mutant strains with reduced c-di-AMP production were cultured under thymidine-limited conditions, and bacterial viability was monitored through intracellular growth assays during infection models. Both oral and intravenous mouse infection experiments were conducted to examine the effect of thymidine availability on bacterial propagation. Additionally, the c-di-AMP-binding protein PstA was genetically deleted to determine its contribution to TLD regulation. These experiments demonstrated that elevated c-di-AMP suppresses bacterial cell death under thymidine starvation.

Cyclic di-AMP for Artificial Metalloribozyme-Catalyzed Enantioselective Diels-Alder Reactions

Qi Q, et al. European Journal of Organic Chemistry, 2020, 2020(8), 4417-4424.

Cyclic di-AMP was utilized as a structural RNA component to construct an artificial metalloribozyme capable of catalyzing enantioselective Diels-Alder reactions. In the experimental design, c-di-AMP was assembled with Cu²⁺ ions to generate a catalytic complex (c-di-AMP·Cu²⁺). Reaction systems were systematically optimized by varying metal cofactors, c-di-AMP/Cu²⁺ molar ratios, buffer systems, and additives. The catalytic activity was evaluated using model Diels-Alder substrates, and product formation was quantified to determine reaction rates and enantiomeric excess. The c-di-AMP·Cu²⁺ complex exhibited up to 80% enantiomeric excess and significantly enhanced reaction kinetics, achieving approximately 300-fold rate acceleration relative to the uncatalyzed reaction.

Cyclic di-AMP for Competitive ELISA-Based Quantification in Bacterial Samples

Underwood AJ, et al. Journal of Microbiological Methods, 2014, 107, 58-62.

Cyclic di-AMP was quantitatively analyzed using a competitive enzyme-linked immunosorbent assay (ELISA) based on a pneumococcal cyclic di-AMP-binding protein (CabP). Bacterial cultures were first grown in Luria-Bertani medium and harvested by centrifugation. Cell pellets were washed with PBS and lysed in Tris-HCl buffer using ultrasonic disruption. The lysates were heated to denature proteins and centrifuged to obtain supernatants containing soluble metabolites. For the ELISA procedure, purified recombinant CabP was immobilized on microtiter plates and blocked with bovine serum albumin. Samples were mixed with biotinylated c-di-AMP and incubated in the coated wells, enabling competitive binding. HRP-conjugated streptavidin was then applied for detection, and color development was measured at 490 nm after sulfuric acid termination. This method enables rapid and sensitive quantification of cyclic di-AMP in bacterial extracts, supporting studies of bacterial signaling pathways.