Class C CpG oligonucleotide - Human / Murine TLR9 ligand

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 are potent inducers of IFN-α from pDC and strong B cell activators [5, 6].

ODN 2395 is a CpG ODN type C specific for human / murine TLR9 [7].

References:

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.
5. Jurk M. et al., 2004. C-Class CpG ODN: sequence requirements and characterization of immunostimulatory activities on mRNA level. Immunobiology. 209(1-2):141-54.
6. Abel K. et al., 2005. Deoxycytidyl-deoxyguanosine oligonucleotide classes A, B, and C induce distinct cytokine gene expression patterns in rhesus monkey peripheral blood mononuclear cells and distinct alpha interferon responses in TLR9-expressing rhesus monkey plasmacytoid dendritic cells. Clin Diagn Lab Immunol. 12(5):606-21.

7. Roda JM. et al., 2005. CpG-containing oligodeoxynucleotides act through TLR9 to enhance the NK cell cytokine response to antibodycoated tumor cells. J Immunol. 175(3):1619-27.

Specifications

Specificity: human/murine TLR9 agonist

Working concentration: 1-5 µM

Solubility:  5 mg/ml in water

ODN 2395 sequence
5’-tcgtcgttttcggcgc:gcgccg-3’ (22 mer)
Note: Bases are phosphorothioate (nuclease resistant), palindrome is underlined.

Quality control:

• Human TLR9 (hTLR9) and murine TLR9 (mTLR9) activity has been tested using HEK-Blue™ hTLR9 and HEK-Blue™ mTLR9 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 2395 is provided lyophilized and is available in three quantities.

tlrl-2395 (formerly tlrl-odnc):

• 200 μg (28.37 nmol) lyophilized ODN 2395

• 1.5 ml sterile endotoxin-free water

tlrl-2395-1 (formerly tlrl-odnc-1):

• 1 mg (141.85 nmol):  tlrl-2395-1 (formerly tlrl-odnc-1)

• 1.5 ml sterile endotoxin-free water

tlrl-2395-5 (formerly tlrl-odnc-5):

• 5 mg (709.25 nmol):  tlrl-2395-5 (formerly tlrl-odnc-5)

• 10 ml sterile endotoxin-free water

 ODN 2395 is shipped at room temperature.

 Upon receipt, store at -20°C.

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