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dc.contributor.authorAllison, Simon J.*
dc.contributor.authorMilner, J.*
dc.date.accessioned2016-10-10T16:02:57Z
dc.date.available2016-10-10T16:02:57Z
dc.date.issued2014
dc.identifier.citationAllison SJ and Milner J (2014) RNA Interference by Single- and Double-stranded siRNA With a DNA Extension Containing a 3' Nuclease-resistant Mini-hairpin Structure. Molecular Therapy - Nucleic Acids. 2: e141.
dc.identifier.urihttp://hdl.handle.net/10454/9834
dc.descriptionYes
dc.description.abstractSelective gene silencing by RNA interference (RNAi) involves double-stranded small interfering RNA (ds siRNA) composed of single-stranded (ss) guide and passenger RNAs. siRNA is recognized and processed by Ago2 and C3PO, endonucleases of the RNA-induced silencing complex (RISC). RISC cleaves passenger RNA, exposing the guide RNA for base-pairing with its homologous mRNA target. Remarkably, the 3′ end of passenger RNA can accommodate a DNA extension of 19-nucleotides without loss of RNAi function. This construct is termed passenger-3′-DNA/ds siRNA and includes a 3′-nuclease-resistant mini-hairpin structure. To test this novel modification further, we have now compared the following constructs: (I) guide-3′-DNA/ds siRNA, (II) passenger-3′-DNA/ds siRNA, (III) guide-3′-DNA/ss siRNA, and (IV) passenger-3′-DNA/ss siRNA. The RNAi target was SIRT1, a cancer-specific survival factor. Constructs I–III each induced selective knock-down of SIRT1 mRNA and protein in both noncancer and cancer cells, accompanied by apoptotic cell death in the cancer cells. Construct IV, which lacks the SIRT1 guide strand, had no effect. Importantly, the 3′-DNA mini-hairpin conferred nuclease resistance to constructs I and II. Resistance required the double-stranded RNA structure since single-stranded guide-3′-DNA/ss siRNA (construct III) was susceptible to serum nucleases with associated loss of RNAi activity. The potential applications of 3′-DNA/siRNA constructs are discussed.
dc.relation.isreferencedbyhttp://dx.doi.org/10.1038/mtna.2013.68
dc.rights(c) 2014 The Authors. This is an Open Access article distributed under the Creative Commons CC-BY license (http://creativecommons.org/licenses/by/3.0/)
dc.subjectAgo2; C3PO; Nuclease-resistant DNA mini-hairpin; RNAi; SIRT1; Therapy; 3'-DNA/siRNA constructs
dc.titleRNA interference by single- and double-stranded siRNA with a DNA extension containing a 3' nuclease-resistant mini-hairpin structure
dc.status.refereedYes
dc.date.Accepted2013-11-06
dc.typeArticle
dc.type.versionPublished version
refterms.dateFOA2018-07-25T13:08:49Z


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