Title: A screen of chemical modifications identifies position-specific modification by UNA to most potently reduce siRNA off-target effects
Authors: Bramsen, Jesper B
Pakula, Malgorzata M
Hansen, Thomas B
Bus, Claus
Langkjær, Niels
Odadzic, Dalibor
Smicius, Romualdas
Wengel, Suzy L
Chattopadhyaya, Jyoti
Engels, Joachim W
Herdewijn, Piet
Wengel, Jesper
Kjems, Jørgen # ×
Issue Date: Sep-2010
Publisher: Oxford University Press
Series Title: Nucleic Acids Research vol:38 issue:17 pages:5761-5773
Abstract: Small interfering RNAs (siRNAs) are now established as the preferred tool to inhibit gene function in mammalian cells yet trigger unintended gene silencing due to their inherent miRNA-like behavior. Such off-target effects are primarily mediated by the sequence-specific interaction between the siRNA seed regions (position 2-8 of either siRNA strand counting from the 5'-end) and complementary sequences in the 3'UTR of (off-) targets. It was previously shown that chemical modification of siRNAs can reduce off-targeting but only very few modifications have been tested leaving more to be identified. Here we developed a luciferase reporter-based assay suitable to monitor siRNA off-targeting in a high throughput manner using stable cell lines. We investigated the impact of chemically modifying single nucleotide positions within the siRNA seed on siRNA function and off-targeting using 10 different types of chemical modifications, three different target sequences and three siRNA concentrations. We found several differently modified siRNAs to exercise reduced off-targeting yet incorporation of the strongly destabilizing unlocked nucleic acid (UNA) modification into position 7 of the siRNA most potently reduced off-targeting for all tested sequences. Notably, such position-specific destabilization of siRNA-target interactions did not significantly reduce siRNA potency and is therefore well suited for future siRNA designs especially for applications in vivo where siRNA concentrations, expectedly, will be low.
ISSN: 0305-1048
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Medicinal Chemistry (Rega Institute)
× corresponding author
# (joint) last author

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