uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
A screen of chemical modifications identifies position-specific modification by UNA to most potently reduce siRNA off-target effects
Show others and affiliations
2010 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 38, no 17, 5761-5773 p.Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2010. Vol. 38, no 17, 5761-5773 p.
National Category
Biological Sciences Chemical Sciences
URN: urn:nbn:se:uu:diva-134673DOI: 10.1093/nar/gkq341ISI: 000282173300019OAI: oai:DiVA.org:uu-134673DiVA: diva2:373665
Available from: 2010-12-01 Created: 2010-11-30 Last updated: 2010-12-01Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full text
By organisation
Bioorganic Chemistry
In the same journal
Nucleic Acids Research
Biological SciencesChemical Sciences

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 194 hits
ReferencesLink to record
Permanent link

Direct link