Imaging proteolytic activity in live cells and animal models.
Galbán, S., Jeon, Y.H., Bowman, B.M., Stevenson, J., Sebolt, K.A., Sharkey, L.M., Lafferty, M., Hoff, B.A., Butler, B.L., Wigdal, S.S., Binkowski, B.F., Otto, P., Zimmerman, K., Vidugiris, G., Encell, L.P., Fan, F., Wood, K.V., Galbán, C.J., Ross, B.D., and Rehemtulla, A.
This paper describes use of GloSenso™ technology to develop a luciferase biosensor for detection of caspase 3/7 activity. The authors stably expressed a caspase-3/7 biosensor (pGloSensor™-30F DEVDG Vector, available as a custom product from Promega) in 1833 breast cells and D54 glioma cells, which were then used to monitor caspase activation in cell culture following addition of the cell-permeable substrate (GloSensor™ cAMP Reagent). The paper demonstrates use of this luciferase biosensor to detect apoptosis in vivo in cultured cells and in a mouse model, and also demonstrates the utility of the technology for high-throughput screening for pharmacologically active compounds. The regulation of apoptosis by caspases is used as an example in this study, biosensors to study other proteases involved in the regulation of cellular processes can be designed using the concepts described in the paper. (4525)
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