ODN TTAGGG (A151)
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TLR9 antagonist - Human preferred
Suppressive oligonucleotide - TLR9, AIM2 and cGAS antagonist
ODN TTAGGG (A151), also known simply as A151, is a synthetic oligonucleotide (ODN) containing 4 repeats of the immunosuppressive TTAGGG motif commonly found in mammalian telomeric DNA .
This ODN was initially identified as a TLR9 antagonist that inhibits immune activation by CpG-containing ODNs . Subsequently, additional targets for this inhibitor were found: by binding to the cytosolic DNA sensors (CDSs) AIM2 and IFI16, A151 prevents AIM2 inflammasome activation [3,4]. This ODN is also described as a cGAS inhibitor, acting through competition with DNA . Overall, these findings show that ODN TTAGGG (A151) is a multiple pattern recognition receptors (PRR) suppressor that can prove useful for immunomodulatory studies.
1. Steinhagen F. et al., 2017. Suppressive oligodeoxynucleotides containing TTAGGG motifs inhibit cGAS activation in human monocytes. Eur J Immunol. DOI: 10.1002/ eji.201747338.
2. Krieg A. et al., 1998. Sequence motifs in adenoviral DNA block immune activation by stimulatory CpG motifs. PNAS 95(21):12631-6.
3. Eichholz K. et al., 2016. Immune-Complexed Adenovirus Induce AIM2-Mediated Pyroptosis in Human Dendritic Cells. PLoS Pathog. 12(9):e1005871.
4. Kaminski J. et al., 2013. Synthetic oligodeoxynucleotides containing suppressive TTAGGG motifs inhibit AIM2 inflammasome activation. J Immunol. 191(7):3876-83.
Dose-dependent inhibition of TLR9 activity in HEK-Blue™ hTLR9 cells. Cells were treated with the human CpG-ODN 2006 (1 μg/ml) in the presence of increasing concentrations of ODN TTAGGG (A151). After overnight incubation, the NF-κB response was determined using QUANTI-Blue™, a SEAP detection reagent. Data represent % inhibition of the signal.
Effect of inflammasome inhibitors on IL-1β release by THP1-HMGB1-Lucia™ cells. Cells were primed with LPS (1 μg/ml) for 3 hours prior to treatment with 200 μg/ml MSU (NLRP3 inflammasome inducer) or 0.5 μg/ml Poly(dA:dT) complexes (AIM2 inflammasome inducer) in the presence of 0.5 μM Z-VAD (pan-capase inhibitor), Ac-YVAD-cmk (caspase-1 inhibitor), MCC950 (NLRP3 inflammasome inhibitor), VX-765 (caspase-1 inhibitor) and ODN TTAGGG (A151; AIM2 inhibitor). After overnight incubation, cell supernatants were added to HEK-Blue™ IL-1β cells for 16 hours. IL-1β levels were determined by assessing SEAP activity in the supernatant using QUANTI-Blue™.
Inhibition of CDS activity in THP1-Dual™ cells. Cells were pre-treated with increasing concentrations of ODN TTAGGG (A151) for 6 hours followed by stimulation with cytosolic double-stranded DNA (1 μg/ml). Prior to experimentation, the DNA was complexed with a transfection reagent to facilitate intracellular delivery. After overnight incubation, the IRF and NF-κB responses were determined using QUANTI-Luc™ and QUANTI-Blue™, respectively.
Working concentration: 100 nM - 10 µM
Solubility: 2.5 mg/ml in water
Molecular weight: 7944 g/mol
ODN TTAGGG sequence
5’- tt agg gtt agg gtt agg gtt agg g -3’ (24 mer)
Note: Bases are phosphorothioate-linked (nuclease resistant).
- Biological activity has been tested using HEK-Blue™ TLR9 cells.
- The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.
ODN TTAGGG (A151) is provided lyophilized and is available in two quantities.
- 200 μg (25.2 nmol) lyophilized ODN TTAGGG (A151)
- 1.5 ml endotoxin-free water
- 1 mg (126 nmol) lyophilized ODN TTAGGG (A151)
- 1.5 ml endotoxin-free water
ODN TTAGGG (A151) is shipped at room temperature.
Upon receipt, store at -20 °C.Back to the top