Human MDA5-KO Dual Reporter A549 Cells

NF-κB-SEAP & IRF-Lucia reporter lung carcinoma

ABOUT

MDA-5 knockout NF-κB-SEAP & IRF-Lucia Reporter Cell Line

A549-Dual™ KO-MDA5 cells were generated from A549-Dual™ cells through the stable knockout of the MDA5 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-MDA5 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-MDA5 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.

Disclaimer:  These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.

SPECIFICATIONS

Specifications

Target

MDA5

Target species

Human

Tested applications

Screening of PRR agonists or inhibitors

Cell type
Epithelial
Growth properties
Adherent
Tissue origin
Human lung carcinoma
Reporter gene
SEAP
Lucia®
Detection method
Colorimetric, Bioluminescence
Antibiotic resistance
Zeocin®
Blasticidin
Growth medium

Complete DMEM (see TDS)

Mycoplasma-free

Verified using Plasmotest

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    A549-Dual™ KO-MDA5 Cells
  • Cat code: 
    a549d-komda5
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml of Zeocin® (100 mg/ml)
  • 1 ml of blasticidin (10 mg/ml)
  • 1 ml of Normocin™ (50 mg/ml)
  • 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution)
  • 1 tube of QUANTI-Luc™ 4 Reagent (sufficient to prepare 25 ml)

Shipping & Storage

  • Shipping method:  Dry ice
  • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

    Caution:

    • Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.

Details

MDA-5 (melanoma-differentiation-associated gene 5, also known as Ifih1 or Helicard) is a cytoplasmic RNA helicase that plays an important role in antiviral response. It senses double-stranded RNA (dsRNA), a replication intermediate for RNA viruses, leading to production of type I interferons (IFNs) in infected cells. MDA-5 and the related RNA helicase RIG-I recognize a complementary set of cytosolic viral dsRNA. However, some viruses such as picornaviruses appear to activate only MDA-5. Interestingly, transfected poly(I:C), a synthetic analog of viral dsRNA, is recognized by both MDA-5 and RIG-I.

DOCUMENTS

Documents

A549-Dual™ KO-MDA5 Cells

Technical Data Sheet

Validation Data Sheet

Safety Data Sheet

Certificate of analysis

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CUSTOMER SERVICE & TECHNICAL SUPPORT

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