5-Vinylphosphonate improves tissue accumulation and efficacy of conjugated siRNAs in vivo
Authors
Haraszti, Reka A.Roux, Loic
Coles, Andrew H.
Turanov, Anton A.
Alterman, Julia F.
Echeverria, Dimas
Godinho, Bruno M. D. C.
Aronin, Neil
Khvorova, Anastasia
UMass Chan Affiliations
Department of MedicineProgram in Molecular Medicine
RNA Therapeutics Institute
Document Type
Journal ArticlePublication Date
2017-07-27Keywords
BiochemistryEnzymes and Coenzymes
Nucleic Acids, Nucleotides, and Nucleosides
Organic Chemicals
Structural Biology
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5-Vinylphosphonate modification of siRNAs protects them from phosphatases, and improves silencing activity. Here, we show that 5-vinylphosphonate confers novel properties to siRNAs. Specifically, 5-vinylphosphonate (i) increases siRNA accumulation in tissues, (ii) extends duration of silencing in multiple organs and (iii) protects siRNAs from 5-to-3 exonucleases. Delivery of conjugated siRNAs requires extensive chemical modifications to achieve stability in vivo. Because chemically modified siRNAs are poor substrates for phosphorylation by kinases, and 5-phosphate is required for loading into RNA-induced silencing complex, the synthetic addition of a 5-phosphate on a fully modified siRNA guide strand is expected to be beneficial. Here, we show that synthetic phosphorylation of fully modified cholesterol-conjugated siRNAs increases their potency and efficacy in vitro, but when delivered systemically to mice, the 5-phosphate is removed within 2 hours. The 5-phosphate mimic 5-(E)-vinylphosphonate stabilizes the 5 end of the guide strand by protecting it from phosphatases and 5-to-3 exonucleases. The improved stability increases guide strand accumulation and retention in tissues, which significantly enhances the efficacy of cholesterol-conjugated siRNAs and the duration of silencing in vivo. Moreover, we show that 5-(E)-vinylphosphonate stabilizes 5 phosphate, thereby enabling systemic delivery to and silencing in kidney and heart.Source
Nucleic Acids Res. 2017 Jul 27;45(13):7581-7592. doi: 10.1093/nar/gkx507. Link to article on publisher's siteDOI
10.1093/nar/gkx507Permanent Link to this Item
http://hdl.handle.net/20.500.14038/40389PubMed ID
28591791Related Resources
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Copyright The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited.Distribution License
http://creativecommons.org/licenses/by-nc/4.0/ae974a485f413a2113503eed53cd6c53
10.1093/nar/gkx507
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Except where otherwise noted, this item's license is described as Copyright The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited.