NF-qB SEAP Reporter Lentivirus

The NF-?B SEAP Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and nondividing cells. The particles contain a SEAP (Secreted Embryonic Alkaline Phosphatase) reporter driven by the NF-?B (nuclear factor kappa-light-chain-enhancer of activated B cells) response element located upstream of the minimal TATA promoter. These lentiviruses also transduce a puromycin selection marker (Figure 1). After transduction, activation of the NF-?B signaling pathway in the target cells can be monitored by measuring SEAP activity. Figure 1. Schematic of the lenti-vector used to generate the NF-?B SEAP Reporter Lentivirus. The role of NF-kappaB (nuclear factor kappa-light chain enhancer of activated B cells) is well-characterized in canonical (classical) and noncanonical (alternative) signaling pathways of inflammation. Two major forms of innate immune sensors are Toll-like receptors (TLR) and NOD/CATERPILLER proteins. Mutations in NOD2 (nucleotide-binding oligomerization domain-containing protein 2) have been linked to chronic autoinflammatory and autoimmune diseases, such as Crohn's disease and Blaus syndrome. Studying the canonical and noncanonical NF-kappaB pathways and the influence of TLR pathways and NOD2 mutations can further our understanding of autoimmune regulation. The use of reporter systems allows for easy assay read outs. SEAP (Secreted Embryonic Alkaline Phosphatase) is a reporter protein that has been extensively used to assess the activity of transcription factors of interest. Since SEAP is secreted, cell lysis is not required to measure reporter activity, simplifying the assay.