In order to monitor NFkB activation in alive cells and facilitate high throughput drug screening, Signosis has developed NFkB secreted luciferase based HEK293 reporter stable cell lines utilizing NFkB DNA binding sequence and TA minimal promoter to drive Gaussia Luciferase expression in response to the corresponding stimuli. Gaussia Luciferase has a small size of 20kDa, over three times smaller than firefly luciferase, and is naturally secreted from the cells. Therefore, the cell line allows for non-invasive monitoring of your signaling pathway of choice—unlike standard reporter stable cell lines that would require lysis of the cells, a small aliquot of cell culture media is all that is needed for reading. This allows for easy and efficient monitoring of the cells in real-time. The cell line has been well validated with dose response and timed course with TNF treatment to confirm the similar responding property to the cell line with firefly luciferase.
 
NFkB Responsive Secreted Luciferase Reporter HEK293 Stable Cell Line is derived from human embryonic kidney, and stably expresses gaussia luciferase reporter gene under the control of the NFkB  response element. This cell line is an ideal cellular model for monitoring the activation of NFkB Receptor Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.
 
The cell line was established by transfection using a pTA-NFkB-Gaussia luciferase reporter vector, which contains 4 repeats of NFkB binding sites, a minimal promoter upstream of the firefly luciferase coding region, along with hygromycin expression vector followed by hygromycin selection. The hygromycin resistant clones were subsequently screened for TNFa-induced luciferase activity.
 
The Gaussia Luciferase is a gene from the copepod Gaussia princeps, which is a small, naturally secreted luciferase of approximately 20kDa. This luciferase is naturally secreted from the cells, which allows for the measurement of reporter activity in the media without harming the cells. Gaussia luciferase requires a coelenterazine-based luciferase substrate for reading and should not be used with firefly luciferase substrate.