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G3-YSD Control

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G3-YSD Control

Control for cGAS Agonist G3-YSD

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200 µg

tlrl-ydnac
+-
$157

No cGAS activation with G3-YSD Control
No cGAS activation with G3-YSD Control

Negative control for G3-YSD

G3-YSD Control is a negative control for G3-YSD, a potent cGAS (cyclic GMP-AMP synthase, cGAMP synthase) agonist. This negative control differs from the G3-YSD agonist in the hairpin-flanking nucleoside trimers. While G3-YSD is flanked with guanosine trimers (G3) conferring its agonist activity, G3-YSD Control is flanked with cytidine trimers (C3) abrogating cGAS activation [1]. cGAS is a critical cytosolic DNA sensor that triggers innate immune responses through the production of type I interferons (IFNs) [1].

More details More details

 

Mode of action:

cGAS detects double-stranded DNA (dsDNA) over 40 bp in length, or stem-loop structures of single-stranded DNA (ssDNA) flanked by unpaired nucleotides [1-3]. Interaction of cGAS with its agonists promotes the synthesis of 2’3’-cGAMP, a second messenger that activates STING (stimulator of interferon genes), and the downstream production of type I interferons (IFNs) and other cytokines [2].
In THP1-Dual™ reporter cells, which express multiple cytosolic DNA sensors including cGAS, intracellular G3-YSD Control (C3-ended Y-form Short DNA) does not activate an interferon regulatory factor (IRF) response compared to intracellular G3-YSD (see figure).

 

Key features of G3-YSD Control:

  • Negative control for G3-YSD
  • Y-form Short DNA flanked with cytidine trimers
  • Each lot is functionally tested

 

Read our review on cGAS.

 

References:

1. Herzner A.M. et al., 2015. Sequence-specific activation of the DNA sensor cGAS by Y-form DNA structures as found in primary HIV-1 cDNA. Nat Immunol. 16(10):1025-33.
2. Li T. & Chen Z.J., 2018. The cGAS-cGAMP-STING pathway connects DNA damage to inflammation, senescence, and cancer. J Exp Med.215(5):1287-1299.
3. Luecke S. et al., 2017. cGAS is activated by DNA in a length-dependent manner. EMBO Rep. 18(10):1707-1715.

Figures

Evaluation of IRF induction
Evaluation of IRF induction

Dose-dependent IRF response in THP1-Dual™ cells. These cells were stimulated with increasing concentrations of G3‑YSD or G3‑YSD Control complexed with LyoVec™, or 1 x 104 U/ml hIFN-β. After overnight incubation, the IRF response was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™ 4 Lucia/Gaussia. For each ligand, the response is expressed relative to that of hIFN-β (taken as 100%).

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Specifications

Activity: Control for G3-YSD (cGAS agonist)

Working concentration: 100 ng - 1 μg/ml

Sequence: 5’ CCC TATATATATGCATATATATA CCC 3’ (26 mer)

Note: G3-YSD Control contains a palindromic sequence for which self-hybridization results in double-stranded DNA. The flanking cytidine trimers in 5' and 3' remain unpaired.

Molecular weight: 7843.3 g/mol

Quality control: 

  • The inability to induce type I interferon has been verified using cellular assays.
  • The absence of bacterial contamination, such as lipoproteins and endotoxins, has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.
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Contents

  • 200 μg G3-YSD Control (C3-ended Y-form Short DNA)
  • 1.5 ml sterile endotoxin-free water

room temperature G3-YSD Control is shipped at room temperature.

store Upon receipt, store at -20°C.

stable Resuspended product is stable for 12 months at -20°C.

Alert Avoid repeated freeze-thaw cycles.

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Details

cGAS

Cyclic GMP-AMP synthase (cGAS, cGAMP synthase) is a critical cytosolic DNA sensor that triggers innate immune responses through the production of type I interferons (IFNs) [1]. In response to cytosolic double‑stranded DNA (dsDNA), cGAS produces the cyclic dinucleotide (CDN) 2’3’-cGAMP.
CDNs bind directly to STING, leading to TBK1‑IRF3-mediated activation of IFN-stimulated response elements (ISRE) in the promoters of IFN-stimulated genes (ISG). The most potent agonist of human STING is 2’3’-cGAMP [2,3].

 

References:

1. Sun L. et al., 2013. Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway. Science 339(6121):786-91.
2. Gao P. et al., 2013. Cyclic [G(2’,5’)pA(3’,5’)p] is the metazoan second messenger produced by DNA-activated cyclic GMP-AMP synthase. Cell. 153(5):1094-107.
3. Ablasser A. et al., 2013. cGAS produces a 2’-5’-linked cyclic dinucleotide second messenger that activates STING. Nature. 498(7454):380-4.

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