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5'ppp-dsRNA Control

5'ppp-dsRNA Control Unit size Cat. code Docs Qty Price
Control for 5'ppp-dsRNA
25 µg
100 µg
tlrl-3prnac
+-
$110.00
5'ppp-dsRNA Control / LyoVec™ Unit size Cat. code Docs Qty Price
Control ligand for RIG-I - complexed with transfection reagent
25 µg
100 µg
tlrl-3prnaclv
+-
$179.00

Negative control for 5′ppp-dsRNA

5’ppp-dsRNA Control is a negative control for 5’ triphosphate double-stranded RNA (5’ppp-dsRNA), 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]. 5’ppp-dsRNA Control is a 19 mer blunt-end dsRNA without a 5’triphosphate.

5’ppp-dsRNA Control must be delivered into the cytoplasm, for example by using a transfection agent, such as LyoVec™.

 

Key features of 5'ppp-dsRNA Control:

  • Negative control for the specific RIG-I specific agonist 5'ppp-dsRNA (it does not contain the 5' triphosphate moiety).
  • Its biological activity has been verified using cellular assays.
  • For your convenience, pre-complexed 5'ppp-dsRNA Control/ 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

RIG-I response to 5'ppp-dsRNA and 5'ppp-dsRNA Control in B16-Blue™ ISG cells
RIG-I response to 5'ppp-dsRNA and 5'ppp-dsRNA Control in B16-Blue™ ISG cells

RIG-I activation with 5’ppp-dsRNA can be easily monitored using B16 Blue™ ISG cells, an IFN regulatory factor (IRF)-inducible secreted embryonic alkaline phosphatase (SEAP) reporter cell line. B16-Blue™ ISG cells respond to stimulation with 5’ppp-dsRNA and Poly(I:C). The 5’ppp-dsRNA Control (non-phosphorylated) does not induce any response in B16-Blue™ ISG cells. Levels of SEAP is determined by colorimetric measurement using QUANTI-Blue™ and reading optical density (O.D.) at 630-655 nm.

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Specifications

Sequence:

5’- GCAUGCGACCUCUGUUUGA -3’ (19 mer)
3’- CGUACGCUGGAGACAAACU -5’ (19 mer)

Formulation: Naked or pre-complexed with LyoVec™

Working Concentration: 300 ng- 1 μg/ml

Solubility:

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

Quality Control:

  • The absence of 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.
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Contents

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

tlrl-3prnac:

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

tlrl-3prnac-100:

  • 100 μg (4x25μg) 5’ppp-dsRNA Control naked.
  • 10 ml sterile endotoxin-free water.

 

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

tlrl-3prnaclv:

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

tlrl-3prnaclv-100:

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

 

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

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

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

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Citations

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