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Title

RNA/aTNA chimeras: RNAi effects and nucleases resistance of single and double stranded RNAs

AuthorsAlagia, Adele ; Eritja Casadellà, Ramón ; Terrazas, Montserrat
Keywords3'-exonuclease
3'-overhang chemical modification
5'-exonuclease
ApoB gene
L-threoninol
RNAi
Serum resistance
Single-stranded siRNA
SiRNA
Issue DateNov-2014
PublisherMultidisciplinary Digital Publishing Institute
CitationMolecules
AbstractThe RNA interference pathway (RNAi) is a specific and powerful biological process, triggered by small non-coding RNA molecules and involved in gene expression regulation. In this work, we explored the possibility of increasing the biological stability of these RNA molecules by replacing their natural ribose ring with an acyclic L-threoninol backbone. In particular, this modification has been incorporated at certain positions of the oligonucleotide strands and its effects on the biological properties of the siRNA have been evaluated. In vitro cellular RNAi assays have demonstrated that the L-threoninol backbone is well tolerated by the RNAi machinery in both double and single-stranded fashion, with activities significantly higher than those evinced by the unmodified RNAs and comparable to the well-known phosphorothioate modification. Additionally, this modification conferred extremely strong resistance to serum and 3′/5′-exonucleases. In view of these results, we applied this modification to the knockdown of a therapeutically relevant human gene such as apolipoprotein B ( ApoB). Further studies on the activation of the innate immune system showed that L-threoninol-modified RNAs are slightly less stimulatory than unmodified RNAs.
Publisher version (URL)DOI: 10.3390/molecules191117872
URIhttp://hdl.handle.net/10261/126493
DOI10.3390/molecules191117872
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