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DNA-Directed RNA Interference: Hairpin Cloning and Expression
Made Easy
Cheryl Bailey, Ph.D., Natalie Betz, Ph.D., Cindy
Sprecher, B.S., Doug Storts, Ph.D., Jolanta Vidugiriene, Ph.D., and
Thomas Yeager, Ph.D.
Promega Corporation
We developed the siSTRIKE™ U6 Hairpin Cloning Systems (Human)
to allow cloning and expression of short hairpin RNAs (shRNAs),
which suppress gene expression in human cells. The short hairpin RNA
target sequence is contained in two complementary, user-supplied,
single-stranded oligonucleotides, which are annealed and ligated
downstream of a U6 promoter in the psiSTRIKE™ Vectors.
Transfection of the resulting vector into cells triggers
transcription of the short hairpin RNA and suppression of the gene
of interest through RNA interference. In this article, we show
efficient suppression of Renilla luciferase and p53 gene
expression using the psiSTRIKE™ Vectors for both transient and
stable transfections.
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