Multiplexing Bioluminescent and Fluorescent Reporters to Monitor Live Cells

Overview

Journal Curr Chem Genomics

Publisher Bentham Open

Specialty Biochemistry

Date 2010 Feb 18

PMID 20161823

Citations 9

Authors

Michael Haugwitz

Omar Nourzaie

Tatiana Garachtchenko

Lanrong Hu

Suvarna Gandlur

Cathy Olsen

Andrew Farmer

Grigoriy Chaga

Hiroaki Sagawa

Affiliations

Soon will be listed here.

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.

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