caa a22 4500 445312513 CHVBK 20180317142635.0 cr unu---uuuuu 170323e20111201xx s 000 0 eng 10.1007/s11095-011-0529-z doi (NATIONALLICENCE)springer-10.1007/s11095-011-0529-z Enhanced Light-Activated RNA Interference Using Phosphorothioate-Based dsRNA Precursors of siRNA [Elektronische Daten] [Ashish Kala, Simon Friedman] ABSTRACT: Purpose: To improve light-activated RNA interference by incorporating phosphorothioate linkages into the dsRNA used. The rationale behind this approach is that the groups have the potential to improve nuclease stability and therefore prevent cleavage of photolabile groups from the RNA termini prior to photolysis. Methods: Photolabile groups (di-methoxy nitro phenyl ethyl or DMNPE) were incorporated into multiple double-stranded precursors of siRNA (dsRNA) that had six, two or no phosphorothioate linkages at the 3′ and 5′ ends of the strands. They were analyzed for their ability to toggle light-activated RNA interference with light and for serum stability. Results: Incorporation of phosphorothioate linkages increased serum stability of all dsRNA examined. Presence of DMNPE groups reduced overall stability of the modified RNA relative to the analogous species without DMNPE modification. DMNPE-modified dsRNA with two phosphorothioate linkages in each strand significantly improved the window of expression toggled by light. Conclusions: Incorporating phosphorothioate groups into dsRNA both stabilizes them towards degradation by serum enzymes, as well as improves them as the basis for light-activated RNA interference. Springer Science+Business Media, LLC, 2011 light-activated gene expression nationallicence phosphorothioate groups nationallicence RNA interference nationallicence serum stability nationallicence siRNA nationallicence Kala Ashish Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri, 2464 Charlotte St., 64108, Kansas City, Missouri, USA aut Friedman Simon Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri, 2464 Charlotte St., 64108, Kansas City, Missouri, USA aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/12(2011-12-01), 3050-3057 0724-8741 28:12<3050 2011 28 11095 https://doi.org/10.1007/s11095-011-0529-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0529-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kala Ashish Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri, 2464 Charlotte St., 64108, Kansas City, Missouri, USA aut NATIONALLICENCE 700 1- Friedman Simon Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri, 2464 Charlotte St., 64108, Kansas City, Missouri, USA aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/12(2011-12-01), 3050-3057 0724-8741 28:12<3050 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer