Current Chemical Genomics and Translational Medicine

2008, 1 : 11-19
Published online 2008 February 25. DOI: 10.2174/1875397300801010011
Publisher ID: CCGTM-1-11

RESEARCH ARTICLE
 Multiplexing Bioluminescent and Fluorescent Reporters to Monitor Live Cells

Michael Haugwitz, *,a , Omar Nourzaie, #,a , Tatiana Garachtchenkoa , Lanrong Hua , Suvarna Gandlura , Cathy Olsenb , Andrew Farmera , Grigoriy Chagaa and Hiroaki Sagawaa
a Clontech Laboratories, Inc., 1290 Terra Bella Ave., Mountain View, CA 94043, USA
b Molecular Devices, now a part of MDS Analytical Technologies, 1311 Orleans Drive, Sunnyvale, CA 94089, USA

* Address correspondence to this author at the Clontech Laboratories, Inc., 1290 Terra Bella Ave., Mountain View, CA 94043, USA; E-mail: Michael_Haugwitz@clontech.com

ABSTRACT

Reporter proteins are valuable tools to monitor promoter activities and characterize signal transduction pathways. Many of the currently available promoter reporters have drawbacks that compromise their performance. Enzyme-based reporter systems using cytosolic luciferases are highly sensitive, but require a cell lysis step that prevents their use in long-term monitoring. By contrast, secreted bioluminescent reporters like Metridia luciferase and Secreted Alkaline Phosphatase can be assayed repeatedly, using supernatant from the same live cell population to produce many sets of data over time. This is crucial for studies with limited amounts of cells, as in the case of stem cells. The use of secreted bioluminescent reporters also enables broader applications to provide more detailed information using live cells; for example, multiplexing with fluorescent proteins. Here, data is presented describing the characteristics of secreted Metridia luciferase and its use in multiplexing applications with either Secreted Alkaline Phosphatase or a fluorescent protein.