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The Use of Bioluminescent Reporter Genes for RNAi Optimization
Jolanta Vidugiriene, Ph.D., Thomas Yeager, Ph.D.,
Brian Almond, Ph.D., Denise Garvin, M.S., Bob Bulleit, Ph.D., and
Doug Storts, Ph.D.
Promega Corporation
The psiCHECK™ Vectors provide a quantitative and rapid approach for the
optimization of RNA interference (RNAi). The vectors enable changes
in expression of a target gene to be determined by monitoring the activity
of a fused reporter gene. In the psiCHECK™ Vectors, Renilla
luciferase is used as the reporter gene. A target gene of interest can be
cloned into the multiple cloning region located downstream of the Renilla
translational stop codon. Initiation of RNAi toward the target gene
results in cleavage and subsequent degradation of the fusion mRNA
including the Renilla luciferase RNA sequence. This RNA degradation
results in decreased Renilla luciferase activity. The decrease in
activity can be monitored and used as a convenient indicator of the RNAi
effect on the target gene. Thus, the psiCHECK™ Vectors allow
quantitative measurements of RNAi activity and can be easily adapted for
use in high-throughput applications.
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