ODN 2007 - TLR9 Agonist

CpG ODN, Class B (Bovine/porcine/chicken)

ABOUT

Class B CpG oligonucleotide - Bovine / porcine / chicken TLR9-preferred ligand

ODN 2007 is a Class B CpG oligonucleotide (ODN). It is a short synthetic single-stranded DNA molecule containing unmethylated CpG dinucleotides (CpG motifs). These unmethylated CpG motifs mimic microbial DNA and act as immunostimulants. ODN 2007 is a ligand of choice for bovine and porcine Toll-like receptor 9 (TLR9) [1-2]. It is also extensively studied as a vaccine adjuvant against the avian virus in chicken [3].

More details More details

 

Mode of action

Stimulatory CpG ODNs are internalized and activate the endosomal receptor TLR9. Activation of TLR9 triggers NF-κB- and interferon regulatory factor (IRF)-mediated pro-inflammatory responses upon the recognition of unmethylated cytosine-phosphorothioate-guanosine (CpG) forms of DNA [4-6]. Unmethylated CpG dinucleotides are a hallmark of microbial (bacterial, viral, fungal, and parasite) DNA, as well as mitochondrial self-DNA [6,7]. Class B (also called Type K) CpG ODNs, such as ODN 2007, contain a full phosphorothioate backbone with one or more CpG dinucleotides. They strongly activate B cells but weakly stimulate IFN-α secretion in plasmacytoid dendritic cells [8]. 

InvivoGen's ODN 2007 is a strong TLR9 agonist verified using our HEK-Blue™ reporter cell lines expressing human or mouse TLR9 (see figure)

 

Key features of ODN 2007

  • Potent activator of TLR9 in cattle, pigs, and chickens.
  • Synthetic ODN with unmethylated CpG motifs
  • Each lot is functionally tested

 

Get more information about CpG ODNs Classes.

Read our review Read our review on TLR9 agonists: double-edged sword for immune therapies.

 

 

References

1. Vordermeier HM, et al., 2005. Synthetic peptide vaccination in cattle: induction of strong cellular immune responses against peptides derived from the Mycobacterium bovis antigen Rv3019c. Vaccine. ;23(35):4375-84.
2. Ouyang K, et al., 2016. Comparative analysis of routes of immunization of a live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine in a heterologous virus challenge study. Vet Res. ;47:45.
3. Raj S, et al., 2023. Treatment with Toll-like Receptor (TLR) Ligands 3 and 21 Prevents Fecal Contact Transmission of Low Pathogenic H9N2 Avian Influenza Virus (AIV) in Chickens. Viruses. ;15(4):977.
4. Kumagai Y. et al., 2008. TLR9 as a key receptor of the recognition of DNA. Adv. Drug. Deliv. Rev. 60(7):795-804.
5. Heinz L.X. et al., 2021. TASL is the SLC15A4-associated adaptor for IRF5 activation by TLR7-9. Nature. 581(7808):316-322.
6. Kayraklioglu N. et al., 2021. CpG oligonucleotides as vaccine adjuvants. DNA Vaccines: Methods and Protocols. Methods in Molecular Biology. Vol. 2197. p51-77.
7. Kumar V., 2021. The trinity of cGAS, TLR9, and ALRs: guardians of the cellular galaxy against host-derived self-DNA. Front. Immunol. 11:624597.
8. Krieg A.M. et al., 1995. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature. 374(6522):546-9.

All products are for research use only, and not for human or veterinary use.

SPECIFICATIONS

Specifications

Source
Synthetic
Target

TLR9

Sequence

5’- tcg tcg ttg tcg ttt tgt cgt t -3’ (22 mer). Note: Bases are phosphorothioate (nuclease resistant).

Molecular weight
7058 g/mol
Working concentration

5 µM (10 μg/ml) for cellular assays

Solubility

5 mg/ml in water

Appearance (form)
Lyophilized
Reconstitution buffer
Sterile endotoxin-free water (provided)
Endotoxin

Negative (tested using EndotoxDetect™ assay)

Tested applications

Cellular assays

Quality control

Each lot is functionally tested and validated using  HEK-Blue™ hTLR9 and  HEK-Blue™ mTLR9 cells.

CONTENTS

Contents

  • Product: 
    ODN 2007
  • Cat code: 
    tlrl-2007-1
  • Quantity: 
    1 mg
Includes:

1.5 ml of sterile endotoxin-free water

Notes:

tlrl-2007-1 (formerly tlrl-podnb-1)

Shipping & Storage

  • Shipping method:  Room temperature
  • Storage:

    • Upon receipt, store product at -20 °C
    Stability: The reconstituted product is stable up to 6 months at -20 °C.

    Caution:

    • Avoid repeated freeze-thaw cycles

Details

CpG ODNs

Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODNs), such as ODN 1018, have been extensively studied as adjuvants [1]. These CpG motifs are present at a 20-fold greater frequency in bacterial DNA compared to mammalian DNA [2]. CpG ODNs are recognized by the Toll-like receptor 9 (TLR9), which is expressed on human B cells and plasmacytoid dendritic cells (pDCs), thereby inducing Th1-dominated immune responses [3]. Pre-clinical studies, conducted in rodents and non-human primates, as well as human clinical trials, have demonstrated that CpG ODNs can significantly improve vaccine-specific antibody responses [1]. Three types of stimulatory CpG ODNs have been identified, types A, B, and C, which differ in their immune-stimulatory activities [4-5]. 

 

Toll-like receptor 9

The Toll-like Receptor 9 (TLR9) is an endosomal receptor that triggers NF-κB- and interferon regulatory factor (IRF)-mediated pro-inflammatory responses upon the recognition of unmethylated cytosine-phosphorothioate-guanosine (CpG) forms of DNA [6-8]. Unmethylated CpG dinucleotides are a hallmark of microbial (bacterial, viral, fungal, and parasite) DNA, as well as mitochondrial self-DNA [8,9]. These TLR9 agonists can be mimicked by synthetic oligonucleotides containing CpG motifs (CpG ODNs), which have been extensively studied to improve adaptive immune responses in the context of vaccination [6,8].

TLR9 is mainly expressed in subsets of Dendritic Cells and B cells of all mammals. In rodents, but not in humans, TLR9 is also expressed in monocytes and macrophages [8]. The structure of the receptor varies by 24% between human TLR9 (hTLR9) and mouse TLR9 (mTLR9) [8]. They recognize different CpG motifs, the optimal sequences being GTCGTT and GACGTT for hTLR9 and mTLR9, respectively [10].
 

 

References:

1. Steinhagen F. et al., 2011. TLR-based immune adjuvants. Vaccine 29(17):3341-55.
2. Hemmi H. et al., 2000. A Toll-like receptor recognizes bacterial DNA. Nature 408:740-5.
3. Coffman RL. et al., 2010. Vaccine adjuvants: Putting innate immunity to work. Immunity 33(4):492-503.
4. 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.
5. 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.
6. Kumagai Y. et al., 2008. TLR9 as a key receptor of the recognition of DNA. Adv. Drug. Deliv. Rev. 60(7):795-804.
7. Heinz L.X. et al., 2021. TASL is the SLC15A4-associated adaptor for IRF5 activation by TLR7-9. Nature. 581(7808):316-322.
8. Kayraklioglu N. et al., 2021. CpG oligonucleotides as vaccine adjuvants. DNA Vaccines: Methods and Protocols. Methods in Molecular Biology. Vol. 2197. p51-77.
9. Kumar V., 2021. The trinity of cGAS, TLR9, and ALRs: guardians of the cellular galaxy against host-derived self-DNA. Front. Immunol. 11:624597.
10. Bauer S. et al., 2001. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proc Natl Acad Sci USA, 98(16):9237-42.

DOCUMENTS

Documents

ODN 2007

Technical Data Sheet

Validation Data Sheet

Safety Data Sheet

Certificate of analysis

Need a CoA ?

CUSTOMER SERVICE & TECHNICAL SUPPORT

Question about this product ?