Wednesday 24 April 2013 at 10:00
Center for Biological Sequence Analysis, meeting room 2, 1 floor in building 101
SiRNAs are the standard tool for efficient gene inactivation in biomedical research and are tested in a growing number of therapeutic applications. However, the major, unresolved shortcoming of siRNA-based gene silencing are sequence-specific off-target effects. Such effects are largely unpredictable because siRNAs can affect partially complementary sequences and function like endogenous microRNAs, which inhibit gene expression on mRNA stability or translational levels. Here we demonstrate that novel, enzymatically generated, siRNA pools- referred to as siPools - containing up to 60 accurately designed siRNAs eliminate off-target effects. This is achieved by the low concentration of each individual siRNA diluting the sequence-specific off-target effects below detection limit. In fact, whole transcriptome analyses reveal that single siRNA transfections can severely affect global gene expression. However, when complex siRNA pools are transfected, only marginal transcriptome alterations are observed. Taken together, we present the enzymatic production of complex but accurately defined siRNA pools with potent on-target silencing but without detectable off-target effects.
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