5'ppp-dsRNA

RIG-I agonist

5’ triphosphate double-stranded RNA (5’ppp-dsRNA) is a synthetic ligand for the retinoic acid-inducible protein I (RIG-I, also known as Ddx58). RIG-I is a cytoplasmic RNA helicase that belongs to the RIG-I-like receptors (RLRs) family and triggers an antiviral immune response by the activation of type-I interferons (IFN-α and -β) [1]. RIG-I detects viral RNAs that exhibit an uncapped 5’-di/triphosphate end and a short blunt-ended double-strand portion, two essential features facilitating discrimination from self-RNAs [2-4].
To achieve stimulation of RIG-I, 5’ppp-dsRNA must be delivered into the cytoplasm, for example by using a transfection agent, such as LyoVec™.

Key features of 5'ppp-dsRNA:

• 5'ppp-dsRNA is a specific RIG-I agonist.
• Its biological activity has been verified using our RLR reporter cell lines.
• For your convenience, pre-complexed 5'ppp-dsRNA/ LyoVec™ is also available.

Read our review on RIG-I and cancer immunotherapy.

Learn more about RIG-I and STING alliance.

References:

1. Yoneyama M. & Fujita T. , 2007. Function of RIG-I-like Receptors in Antiviral Innate Immunity. J. Biol. Chem. 282:15315-8.
2. Schlee M. et al., 2009. Recognition of 5’ triphosphate by RIG-I helicase requires short blunt double-stranded RNA as contained in pandhandle of negative-strand virus. Immunity. 17;31(1):25-34.
3. Schmidt A. et al., 2009. 5’-triphosphate RNA requires base-paired structures to activate antiviral signaling via RIG-I. PNAS 106(29):12067-72.
4. Schlee M. & Hartmann G., 2010. The chase for the RIG-I ligand - recent advances. Mol Ther. 18(7):1254-62.

Figures

A549-Dual™ cell stimulation with 3p-hpRNA induces higher ISG and NF-κB responses than with 5’ppp-dsRNA.
Dose responses of A549-derived cells stimulated with 0.1-100 ng/ml 3p-hpRNA (complexed to LyoVec™) or 0.3 ng/ml to 1 μg/ml 5’ppp-dsRNA (complexed to LyoVec™). After overnight incubation, the ISG response was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™ 4 Lucia/Gaussia and expressed as relative light units (RLUs) (a), and the NF-κB response was determined using QUANTI-Blue™ Solution, a SEAP detection reagent, and by reading the optical density (OD) at 655 nm (b).

RAW-Lucia™ and HEK-Lucia™ RIG-I cell stimulation with 3p-hpRNA induces a better ISG response than with 5’ppp-dsRNA.

RAW-derived (a) and HEK-derived (b) cells were stimulated with 1 μg/ml 5’ppp-dsRNA (complexed to LyoVec™) or 3p-hpRNA (complexed to LyoVec™).
After overnight incubation, the ISG response was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™ 4 Lucia/Gaussia and expressed as relative light units (RLUs).

Specifications

Specificity: RIG-I agonist

Sequence:
5’- pppGCAUGCGACCUCUGUUUGA -3’ (19 mer)
         3’- CGUACGCUGGAGACAAACU -5’ (19 mer)

Formulation: Naked or pre-complexed with LyoVec™

Working Concentrations:

• 300 ng- 1 μg/ml for Naked 5'ppp-dsRNA
• ​1-10 µg/ml for pre-complexed 5'ppp-dsRNA/LyoVec™

Solubility:

• 0.1 mg/ml in water for Naked 5'ppp-dsRNA
• 50 µg/ml in water for pre-complexed 5'ppp-dsRNA/LyoVec™

Quality Control:

• The biological activity has been verified using cellular assays.
• The absence of bacterial contamination (lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.

Contents

5’ppp-dsRNA Naked is available in two quantities:

tlrl-3prna:

• 25 μg 5’ppp-dsRNA naked.
• 2 ml sterile endotoxin-free water.

tlrl-3prna-100:

• 4 x 25 μg 5’ppp-dsRNA naked.
• 10 ml sterile endotoxin-free water.

5’ppp-dsRNA/LyoVec™ is available in two quantities:

tlrl-3prnalv:

• 25 μg 5’ppp-dsRNA complexed with 50 µg of LyoVec™
• 2 ml sterile endotoxin-free water

tlrl-3prnalv-100:

• 4 x 25 μg 5’ppp-dsRNA complexed with 50 µg of LyoVec™
• 10 ml sterile endotoxin-free water

5’ppp-dsRNA and  5’ppp-dsRNA/LyoVec™ are provided lyophilized and shipped at room temperature.

Store lyophilized 5’ppp-dsRNA and  5’ppp-dsRNA/LyoVec™ at -20°C.

Lyophilized product is stable for 1 year when stored -20°C.