Class A CpG oligonucleotide - Human TLR9 ligand

CpG oligonucleotides (ODNs) are synthetic ODNs that contain unmethylated CpG dinucleotides in particular sequence contexts (CpG motifs).

These CpG motifs are present at a 20-fold greater frequency in bacterial DNA compared to mammalian DNA. CpG ODNs activate Toll-like receptor 9 (TLR9), leading to strong immunostimulatory effects.

ODN 2216 is a Class-A CpG ODN that preferentially binds to human TLR9.

Class A CpG ODNs are characterized by a phosphodiester central CpG-containing palindromic motif and a phosphorothioate 3’ poly-G string.

They induce high IFN-α production in plasmacytoid dendritic cells (pDC) but are weak stimulators of TLR9-dependent NF-κB signaling.

Specifications

Specificity: human TLR9 agonist.

Working concentration: 1-5 µM.

Solubility:  5 mg/ml in water.

ODN 2216 sequence:
5’-ggGGGACGA:TCGTCgggggg-3’ (20 mer).
Note: Bases shown in capital letters are phosphodiester, and those in lower case are phosphorothioate (nuclease resistant). Palindrome is underlined.

Quality control:

• TLR9 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.

Contents

ODN 2216 is provided lyophilized and is available in three quantities.

tlrl-2216 (formerly tlrl-hodna):

• 200 μg (31 nmol) lyophilized ODN 2216

• 1.5 ml sterile endotoxin-free water

tlrl-2216-1 (formerly tlrl-hodna-1):

• 1 mg (155 nmol) lyophilized ODN 2216

• 1.5 ml sterile endotoxin-free water

tlrl-2216-5 (formerly tlrl-hodna-5):

• 5 mg (777 nmol) lyophilized ODN 2216

• 10 ml sterile endotoxin-free water

 ODN 2216 is shipped at room temperature.

 Upon receipt, store at -20?°C.

Description

CpG ODNs are synthetic oligonucleotides that contain unmethylated CpG dinucleotides in particular sequence contexts (CpG motifs) [1]. These CpG motifs are present at a 20-fold greater frequency in bacterial DNA compared to mammalian DNA. CpG ODNs are recognized by Toll-like receptor 9 (TLR9) leading to strong immunostimulatory effects [2].

Three types of stimulatory CpG ODNs have been identified, types A, B and C, which differ in their immune-stimulatory activities [3-4]. Type A CpG ODNs are characterized by a phosphodiester central CpG-containing palindromic motif and a phosphorothioate 3’ poly-G string. They induce high IFN-α production from plasmacytoid dendritic cells (pDC) but are weak stimulators of TLR9-dependent NF-κB signaling. Type B CpG ODNs contain a full phosphorothioate backbone with one or more CpG dinucleotides. They strongly activate B cells but stimulate weakly IFN-α secretion. Type C CpG ODNs combine features of both types A and B. They contain a complete phosphorothioate backbone and a CpG-containing palindromic motif. Type C CpG ODNs induce strong IFN-α production from pDC and B cell stimulation.

ODN 2216 is a CpG ODN type A with a preference for human TLR9.

1. Krieg, A.M. et al., 1995. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature, 374(6522):546-9.
2. Bauer, S. et al., 2001. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. PNAS. 98(16):9237-42.
3. Krug A. et al., 2001. Identification of CpG oligonucleotide sequences with high induction of IFN- alpha/beta in plasmacytoid dendritic cells. Eur J Immunol, 31(7): 2154-63.
4. Marshall JD. et al., 2005. Superior activity of the type C class of ISS in vitro and in vivo across multiple species. DNA Cell Biol. 24(2):63-72.

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