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5’-cctcaatagggtgagggg-3’ (lower case letters: phosphorothioate linkage: nuclease resistant)
Reconstitution
For a 100μM stock solution, dissolve total vial content in 1700μl (1mg size) or 170μl (100μg size) sterile endotoxin-free water or PBS.
Formulation
Lyophilized. Sterile.
Biological Activity
Particularly potent sequence of an inhibitory ODN for in vivo use in rodents (50-150µg per injection): based on class I (ODN H154) and class II (ODN A151) sequences to combine inhibitory activity on TLRs and STAT signaling. Negative control: Neutral-ODN (Cat. No.: IAX-200-202).
Endotoxin Content
<0.002EU/μg
Declaration
Manufactured by Innaxon.
Other Product Data
Contains: 100μg size includes 1.5ml ddWater Endotoxin-free (sterile) (Cat. No.: IAX-900-002-LD15). 1mg size includes 10ml ddWater Endotoxin-free (sterile) (Cat. No.: IAX-900-002-L010). Reconstitution Note: Add 50% of solvent and let dissolve for 10min. Add remaining 50% of the solvent and mix thoroughly. Moderate warming may aid dissolving.
Avoid freeze/thaw cycles. After reconstitution, prepare aliquots and keep aqueous stock solutions for 1 day at 4°C or store at -20°C (shelf-life 6 months).
Use/Stability
Stable for at least 2 years after receipt when stored at +4°C.
In recent years several groups have studied the sequence requirements, specificity, signaling pathways and kinetics of the TLR (Toll-like receptor) 9 suppression by inhibitory oligonucleotide motifs, which led to a class of novel inhibitory oligonucleotide (iODNs), that is independent of the previously thought species preference. Subsequently it has been discovered that telomeric DNA repeats (TTAGGG)n can block immune activation by CpG-ODNs. Short, 11-15 base long oligonucleotides were synthesized that were capable of potently inhibiting CpG-stimulation. The optimal inhibitory DNA motif consists of a pyrimidine-rich triplet, preferably CCT, which is positioned 5- to the GGG sequence in a singlestranded DNA molecule. Additionally, both the optimal spacing between the CCT and GGG motifs, as well as their relative order to each other, is of crucial importance for the inhibitory DNA action. Interestingly, although both TLR7/TLR8 ligands and bacterial DNA share the endosomal compartment for receptor binding and signal transduction, certain iODNs (G-type) suppress only TLR9-mediated activation, whereas prototype class I iODN may also interfere with the activation via the TLR7/TLR8 pathway. Recently, intriguing evidence has been presented that for some iODN classes the immuno-modulatory biological activity shows only limited sequence dependency or may not even involve TLR-mediated uptake and signaling pathways. For example iODNs of the class II are thought to act on immune activation through inhibition of STAT signaling and independent of TLR signaling via binding to a yet to be identified 'ODN-receptor'. Slightly modified phosphodiester versions of the most potent inhibitory ODNs were also able to profoundly block the immune activation of macrophages and just recently prove to be valuable tools for in vivo use in experimental animal models of inflammatory and auto-immune diseases. Based upon these recent insights the following classification for iODNs has been suggested:
Class I: G-stretch ODNs: TLR9-specific competitors, some iODNs may also affect TLR7 and TLR8 signaling Class II: ODNs with telomeric repeats: TLR-independent inhibitors of STAT signaling (cellular uptake via an 'ODN receptor'?) Class III: Inhibitors of DNA uptake in a sequence independent manner Class IV: Long phosphorothioate ODNs as direct competitors of TLR9 signaling in a sequence independent manner
Product References
DNA Motifs suppressing TLR9 responses: A. Trieu, et al.; Crit. Rev. Immunol. 26, 527 (2006)
Inhibitory oligodeoxynucleotides-therapeutic promise for systemic autoimmune diseases? P. Lenert; Clin. Exp. Immunol. 140, 1 (2005)
Immunotherapeutic utility of stimulatory and suppressive oligodeoxynucleotides: K.J. Ishii, et al.; Curr. Opin. Mol. Ther. 6, 166 (2004)
Suppressive oligodeoxynucleotides protect mice from lethal endotoxic shock: H. Shirota, et al.; J. Immunol. 174, 4579 (2005)
Suppressive oligodeoxynucleotides inhibit CpG-induced inflammation of the mouse lung: H. Yamada, et al.; Crit. Care Med. 32, 2045 (2004)
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