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Blood, Vol. 96 No. 1 (July 1), 2000:
pp. 321-326
Novel in vitro assay for the detection of pharmacologic
inducers of fetal hemoglobin
Evangelia Skarpidi,
George Vassilopoulos,
Qiliang Li, and
George Stamatoyannopoulos
From the Division of Medical Genetics, University of Washington,
Seattle, WA.
Current techniques for identifying fetal hemoglobin (HbF) inducers
are complex and time consuming. We developed a rapid and efficient
method for detecting HbF inducers. Our system uses a recombinant DNA
construct in which the coding sequences of 2 different luciferase
reporter genes, firefly and renilla, are substituted for those of human
 and  globin genes, respectively. The activity of these genes
can be distinguished by a simple, highly sensitive enzymatic assay in
cell lysates. GM979 cells stably transfected with the construct are
cultured in the presence of compounds, and their effects are determined
by measuring the changes in activity of the 2 luciferase genes.
Specific globin gene inducers are recognized by their ability to
increase -firefly luciferase (F) gene activity
significantly more than -renilla luciferase (R) gene
activity, identified by an increased ratio of -firefly luciferase
activity over total luciferase activity. These results suggest that the
use of the 2 luciferase reporter genes provides a simple, highly
sensitive, and reproducible system for the detection of compounds that
increase -globin gene expression. It can therefore be used for the
screening of chemical agents that may have -globin gene inducibility.
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