Cre Stoplight: A Red/Green Fluorescent Reporter of Cre Recombinase Expression in Living Cells
Author(s) -
Yvonne S. Yang,
Thomas E. Hughes
Publication year - 2001
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/01315st03
Subject(s) - cre recombinase , green fluorescent protein , plasmid , transfection , microbiology and biotechnology , biology , recombinase , reporter gene , cre lox recombination , gene , genetics , transgene , gene expression , recombination , genetically modified mouse
The Cre/lox system is a powerful genetic tool with which to manipulate the genome. Here, we describe the development of a simple reporter system for Cre recombinase, called the Cre Stoplight. In the absence of Cre, the red fluorescent protein is expressed; when Cre catalyzes a recombination event, the green fluorescent protein is produced. Testing this system in transiently transfected cells showed that it produced robust signals (90% of the cells converted from red to green) when equal amounts of the plasmids encoding Cre recombinase and the Cre Stoplight were used. A 1:100 ratio of enzyme to reporter plasmid produced similar results, and a 1:10000 ratio was necessary to significantly reduce the number of cells converting to green (1%).
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