title: Rational design of therapeutic siRNAs: minimizing off-targeting potential to improve the safety of RNAi therapy for Huntington's disease.
abstract: RNA interference (RNAi) provides an approach for the treatment of many human diseases. However, the safety of RNAi-based therapies can be hampered by the ability of small inhibitory RNAs (siRNAs) to bind to unintended mRNAs and reduce their expression, an effect known as off-target gene silencing. Off-targeting primarily occurs when the seed region (nucleotides 2-8 of the small RNA) pairs with sequences in 3'-UTRs of unintended mRNAs and directs translational repression and destabilization of those transcripts. To date, most therapeutic RNAi sequences are selected primarily for gene silencing efficacy, and later evaluated for safety. Here, in designing siRNAs to treat Huntington's disease (HD), a dominant neurodegenerative disorder, we prioritized selection of sequences with minimal off-targeting potentials (i.e., those with a scarcity of seed complements within all known human 3'-UTRs). We identified new promising therapeutic candidate sequences which show potent silencing in cell culture and mouse brain. Furthermore, we present microarray data demonstrating that off-targeting is significantly minimized by using siRNAs that contain "safe" seeds, an important strategy to consider during preclinical development of RNAi-based therapeutics.
authors.affiliation: Department of Internal Medicine, University of Iowa, Iowa City, Iowa 52242, USA.
authors.forename: Ryan L
authors.initials: RL
authors.lastname: Boudreau
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Spengler
authors.forename: Beverly L
authors.initials: BL
authors.lastname: Davidson
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism / Q000473:pathology; D003342:Corpus Striatum / Q000378:metabolism / Q000473:pathology; D000229:Dependovirus / Q000235:genetics; D015195:Drug Design; D020868:Gene Silencing; D018390:Gene Targeting; D006801:Humans; D006816:Huntington Disease / Q000235:genetics* / Q000628:therapy*; D007124:Immunoenzyme Techniques; D051379:Mice; D035683:MicroRNAs / Q000008:administration & dosage / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D050486:Serotonin Plasma Membrane Transport Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: D035683:MicroRNAs; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D050486:Serotonin Plasma Membrane Transport Proteins; C493231:Slc6a4 protein, mouse