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A549-Dual™ KO-MAVS Cells

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A549-Dual™ KO-MAVS cells

MAVS knockout NF-kB-SEAP & IRF-Lucia reporter lung carcinoma cells

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3-7 x 10e6 cells

a549d-komavs
+-
$1,752

MAVS knockout NF-kB-SEAP & IRF-Lucia Reporter Cell Line

A549-Dual™ KO-MAVS cells were generated from A549-Dual™ cells through the stable knockout of the MAVS gene. They are adherent epithelial cells derived from the human A549 lung carcinoma cell line by stable integration of two inducible reporter constructs.

The A549 cell line, a cellular model for asthma and respiratory infections, expresses many pattern recognition receptors (PRRs), including RIG-I [1,2], and the Toll-like receptors (TLRs) TLR2 [3], TLR3, and TLR5 but not TLR4 [3].

A549-Dual™ KO-MAVS and A549-Dual™ cells express a secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of the IFN-β minimal promoter fused to five NF-κB binding sites.

They also express the secreted Lucia luciferase reporter gene under the control of an ISG54 minimal promoter in conjunction with five IFN-stimulated response elements.

As a result, they allow to simultaneously study the NF-κB pathway, by assessing the activity of SEAP, and the interferon regulatory factor (IRF) pathway, by monitoring the activity of Lucia luciferase. Both reporter proteins are readily measurable in the cell culture supernatant when using QUANTI-Blue™ Solution, a SEAP detection reagent, and QUANTI-Luc™ 4 Lucia/Gaussia, a Lucia and Gaussia luciferase detection reagent. 

A549-Dual™ KO-MAVS cells are resistant to blasticidin and Zeocin®.

 

References:

1. Kolokoltsova OA. et al., 2014. RIG-I enhanced interferon independent apoptosis upon Junin virus infection. PLoS One. 9:e99610.
2. Hagmann CA. et al., 2013. RIG-I detects triphosphorylated RNA of Listeria monocytogenes during infection in non-immune cells. PLoS One. 8:e62872.
3. Slevogt H. et al., 2007. Moraxella catarrhalis is internalized in respiratory epithelial cells by a trigger-like mechanism and initiates a TLR2- and partly NOD1-dependent inflammatory immune response. Cell Microbiol. 9(3):694-707.

Figures

IRF INDUCTION (Lucia luciferase reporter)
IRF INDUCTION (Lucia luciferase reporter)

A549-Dual™ (parental cell line) and A549‑Dual™ KO‑MAVS cells were stimulated with hIFN-α (1 x 104 U/ml), 5’ppp-dsRNA/LyoVec™ (1 μg/ml), inactivated NDV (5 x 105 U/ml), poly(I:C) HMW/LyoVec™ (100 ng/ml), poly(dA:dT)/LyoVec™ (10 ng/ml) and poly(I:C) HMW (1 μg/ml). After a 24h incubation, IRF activation was determined by measuring the relative light units (RLUs) in a luminometer using QUANTI‑Luc™, a Lucia luciferase detection reagent. The IRF induction of each ligand is expressed relative to that of hIFN-α at 1 x 104 U/ml (taken as 100%).

NF-κB INDUCTION (SEAP reporter)
NF-κB INDUCTION (SEAP reporter)

A549-Dual™ and A549‑Dual™ KO-MAVS cells were incubated with 5’ppp-dsRNA /LyoVec™ (1 μg/ml), NDV (5 x 106 U/ml), poly(I:C) HMW/LyoVec™ (100 ng/ml), and poly(I:C) HMW (1 μg/ml). After a 24h incubation, NF‑kB activation was determined using QUANTI‑Blue™, a SEAP detection reagent, and by reading the optical density (OD) at 655 nm.

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Specifications

Antibiotic resistance: Zeocin®blasticidin

Growth Medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) fetal bovine serum (FBS), 100 U/ml penicillin, 100 µg/ml streptomycin, 100 µg/ml Normocin™

Freezing Medium: DMEM with 20% FBS and 10% (v/v) DMSO

Test Medium for use with QUANTI-Blue™: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated FBS (30 min at 56°C), 100 U/ml penicillin, 100 µg/ml streptomycin

Quality control:

  • MAVS knockout has been verified by functional assays and DNA sequencing.
  • The stability of this cell line for 20 passages following thawing has been verified.
  • A549-Dual™ KO-MAVS cells are guaranteed mycoplasma-free.

 

These products are covered by a Limited Use License (See Terms and Conditions).

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Contents

Shipped on dry iceShipped on dry ice (Europe, USA, Canada, and some areas in Asia)

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Description

Mitochondrial antiviral-signaling protein (MAVS; also known as IPS‑1, CARDIF, VISA) is an adaptor protein that plays a critical role in the immune response to viral infection. The innate immune system senses intracellular double-stranded RNA (dsRNA), a replication intermediate for RNA viruses, through two RNA helicases: retinoic acid inducible gene-I (RIG-I) and melanoma differentiation-association gene 5 (MDA5). These two sensors recognize different ligands, yet both signal through MAVS. Specifically, upon recognition of dsRNA, they are recruited by MAVS to the outer membrane of the mitochondria leading to the activation of interferon regulatory factor 3 (IRF3), which in turn regulates the expression of type I interferons (IFNs).

A549-Dual KO MAVS pathway

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FAQ Cell Lines

Visit our FAQ Any questions about our cell lines ? Visit our frequently asked questions page

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Notification:  This product is for internal research use only. Additional rights may be available. Please visit InvivoGen’s Terms and Conditions.

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