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BMP/SMAD1/5/8
Principle
Bone morphogenic protein (BMP) is involved in embryogenesis, development of many organ systems and adult tissue homeostasis. Smads (Smad1/5/8) are activated during the signal transduction and then form a complex with Smad4, translocate into the nucleus where they regulate expression of transcriptional factors and transcriptional coactivators. Deficiency in BMP production or functionality usually leads to marked defects or severe human diseases associated with most organ systems. Monitoring the BMP activity is essential to research of the BMP signaling-associated diseases and conduct drug discovery.
Signosis has established BMP luciferase reporter Hek293 stable cell line that has been stably transfected with pTA-BMP-luciferase reporter vector, which contains 4 repeats of BMP binding sites, a minimal promoter upstream of the firefly luciferase coding region. Therefore, the cell line can be used as a reporter system for monitoring the activation of BMP triggered by stimuli treatment, enforced gene expression and gene knockdown.
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TF luciferase reporter stable cell line utilizes artificial promoter constructs to drive luciferase expression. The promoter region can consists of multiple repeats of a cis-element TF binding site, a DNA fragment from the promoter region of a known TF downstream gene, or a DNA fragment containing putative/known TF binding sites. There are several ways that a TF can be activated, such as through extracellular stimuli or through intracellular signaling pathways. Once activated, the TF translocates to the nucleus and often interacts with relevant co-factors to drive gene expression. Once luciferase is expressed, it can generate light in an enzymatic assay and the amount of light measured is positively correlated with the level of TF activation.