5'ppp-dsRNA/LyoVec™
5'ppp-dsRNA/LyoVec™ | Unit size | Cat. code | Docs | Qty | Price |
---|---|---|---|---|---|
RIG-I Agonist - complexed with transfection reagent |
25 µg 100 µg |
tlrl-3prnalv |
RIG-I Agonist - complexed with transfection reagent
5’ triphosphate double stranded RNA (5’ ppp-dsRNA) is a synthetic ligand for retinoic acid-inducible protein I (RIG-I).
RIG-I is a cytosolic pattern recognition receptor that senses pathogen-associated molecular patterns (PAMPs) on viral RNA and triggers an antiviral immune response by the activation of type-I interferons (IFN-α and -β) [1].
RIG-I shares the identification of the dsRNA structure with another sensor MDA-5 (recognition of poly(I:C)/LyoVec™) and is specifically activated by the uncapped 5’ triphosphate moiety on viral RNA [2].
5’ ppp-dsRNA is complexed with the cationic lipid LyoVec™ to facilitate its uptake.
References:
1. Yoneyama M. & Fujita T., 2007. Function of RIG-I-like Receptors in Antiviral Innate Immunity. J. Biol. Chem. 282: 15315-8.
2. Hornung V. et al., 2006. 5'-Triphosphate RNA is the ligand for RIG-I. Science. 314(5801):994-7.
Specifications
Specificity: RIG-I agonist
Working Concentration: 300 ng- 1 μg/ml
Endotoxin level: EndoFit™ (<0.001 EU/µg)
Solubility: 0.05 mg/ml in water
Formulation: complexed with the transfection reagent LyoVec™
5’ppp-dsrna
5’- pppGCAUGCGACCUCUGUUUGA -3’ (19 mer)
3’- CGUACGCUGGAGACAAACU -5’ (19 mer)
Contents
5’ppp-dsRNA/LyoVec™ is provided lyophilized.
This product is available in two sizes:
- tlrl-3prnalv :
- 25 μg 5’ppp-dsRNA/LyoVec™.
- 1.5 ml sterile endotoxin-free water.
- tlrl-3prnalv-100:
- 100 μg (4x25μg) 5’ppp-dsRNA/LyoVec™.
- 10 ml sterile endotoxin-free water.
Note: Each vial contains 25 μg of 5’ppp-dsRNA complexed with 50 μg LyoVec™.
5’ppp-dsRNA/LyoVec™ is shipped at room temperature.
Store lyophilized 5’ppp-dsRNA/Lyovec at -20°C.
Lyophilized product is stable for 1 year when stored -20°C.
Description
Induction of RIG-I with 5’ppp-dsRNA/LyoVec™ can be studied in a variety of cells expressing RIG-I.
If your cell line does not naturally express the RIG-I gene, transfect with a plasmid expressing a RIG-I gene, such as pUNO-hRIG-I or pUNO-mRIG-I.
RIG-I activation can be easily monitored using B16-Blue™ ISG cells, an IFN Regulatory Factor (IRF)-Inducible SEAP reporter cell line. In these cells, activation of RIG-I triggers the secretion of IFNs that results in the production of SEAP.
A protocol for the induction of RIG-I using B16-Blue™ ISG cells is given below:
- - Resuspend 5’ppp-dsRNA/LyoVec™ in water.
- - Stimulate cells with 1 - 10 µg/ml 5’ppp-dsRNA/LyoVec™ for 16 - 24 hours.
Note: The use of 5’ppp-dsRNA Control (as a negative control) and Poly(I:C)/LyoVec™ (as a positive control) is highly recommended.
- - Monitor induction of RIG-I by measuring the levels of SEAP in the cell culture supernatant using QUANTI-Blue™, a SEAP detection reagent.