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KO-MDA5 dual reporter monocytes for RLR pathway studies

Reporter systems in THP1-Dual™-derived cells

InvivoGen offers a new series of THP1-Dual™ cell lines, derived from the human THP-1 monocytic cell line, and specifically designed for assessing the role of RIG-I, MDA5, and MAVS in the cytosolic RNA sensing pathways.

THP1-Dual™ KO-MDA5 cells express two inducible reporter genes, allowing the concomitant study of the IRF and NF-κB pathways, by monitoring the Lucia luciferase and SEAP (secreted embryonic alkaline phosphatase) activities, respectively. In addition, these cells feature a stable knockout of the MDA5 ((Melanoma Differentiation Associated gene 5) gene.

RIG-I and MDA5 are two distinct sensors of viral double-stranded RNA (dsRNA), a replication intermediate for RNA viruses [1-3]. Upon recognition of dsRNA, RIG-I and MDA5 are recruited by the MAVS adaptor to the outer membrane of the mitochondria leading to the activation of several transcription factors, including interferon regulatory factors (IRFs) and NF-κB [3].

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KEY FEATURES

Verified knockout of the MDA5 gene (PCR, DNA sequencing, Western blot, and functional assays)
Functionally validated with a selection of PRR ligands and cytokines
Readily assessable Lucia luciferase and SEAP reporter activities
The stability for 20 passages, following thawing, has been verified
Guaranteed mycoplasma-free

APPLICATIONS

Defining the role of MDA5 in cytosolic RNA sensing pathways
Highlighting possible overlap between RIG-I/MDA5 and STING signaling functions
Developing novel specific inhibitors of the RLR signaling pathway

References

1. Gebhardt A. et al., 2017. Discrimination of Self and Non-Self Ribonucleic Acids. Journal of Interferon & Cytokine Research 37: 184-97.
2. Pichlmair A. et al., 2006. RIG-I mediated antiviral responses to single-stranded RNA bearing 5’-phosphates. Science 314:997-1001.
3. Kawai T. et al., 2005. IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat Immunol. 6(10):981-988.

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