Mouse MDA5-KO Reporter RAW 264.7 Cells

MDA-5 knockout IRF-inducible Lucia luciferase reporter mouse macrophages

RAW-Lucia^™ ISG-KO-MDA5 cells were generated from RAW-Lucia^™ ISG cells through the stable knockout of the MDA-5 gene. These cells derive from the murine RAW 264.7 macrophage cell line, which has been reported to express many pattern recognition receptors (PRRs), including the dsRNA sensors MDA-5 [1] and RIG-I [2,3] along with their adaptor protein IPS-1 (also known as MAVS) [3].

RAW-Lucia^™ ISG-KO-MDA5 and RAW-Lucia^™ ISG cells can be used to study the role of MDA-5 in a mouse macrophage cell line by monitoring of interferon regulatory factor (IRF)-induced Lucia luciferase activity. They express the gene for secreted Lucia luciferase under the control of the I-ISG54 promoter, which is comprised of the IFN-inducible ISG54 promoter enhanced by a multimeric ISRE. The levels of IRF-induced Lucia in the cell culture supernatant can be easily monitored using QUANTI-Luc^™ 4 Lucia/Gaussia, a Lucia and Gaussia luciferase detection reagent.

RAW-Lucia^™ ISG-KO-MDA5 cells are resistant to Zeocin®.

References:

1. Hasan M. et al., 2011. Antimicrobial peptides inhibit polyinosinic-polycytidylic acid-induced immune responses. J Immunol. 187(11):5653-9.
2. Melchjorsen J. et al., 2005. Activation of innate defense against a paramyxovirus is mediated by RIG-I and TLR7 and TLR8 in a cell-type-specific manner. J Virol. 79:12944-51.
3. Yamashita M. et al., 2013. Antiviral innate immunity disturbs podocyte cell function. J Innate Immun. 5:231-41.