Abasic pivot substitution harnesses target specificity of RNA interference
- Authors
- Lee, Hye-Sook; Seok, Heeyoung; Lee, Dong Ha; Ham, Juyoung; Lee, Wooje; Youm, Emilia Moonkyung; Yoo, Jin Seon; Lee, Yong-Seung; Jang, Eun-Sook; Chi, Sung Wook
- Issue Date
- 12월-2015
- Publisher
- NATURE PUBLISHING GROUP
- Keywords
- Abasic pivot substitution; Ago; miRNA; Off-target effect; siRNA; Spacer
- Citation
- NATURE COMMUNICATIONS, v.6
- Indexed
- SCIE
SCOPUS
- Journal Title
- NATURE COMMUNICATIONS
- Volume
- 6
- URI
- https://scholar.korea.ac.kr/handle/2021.sw.korea/91646
- DOI
- 10.1038/ncomms10154
- ISSN
- 2041-1723
- Abstract
- Gene silencing via RNA interference inadvertently represses hundreds of off-target transcripts. Because small interfering RNAs (siRNAs) can function as microRNAs, avoiding miRNA-like off-target repression is a major challenge. Functional miRNA-target interactions are known to pre-require transitional nucleation, base pairs from position 2 to the pivot (position 6). Here, by substituting nucleotide in pivot with abasic spacers, which prevent base pairing and alleviate steric hindrance, we eliminate miRNA-like off-target repression while preserving on-target activity at similar to 80-100%. Specifically, miR-124 containing dSpacer pivot substitution (6pi) loses seed-mediated transcriptome-wide target interactions, repression activity and biological function, whereas other conventional modifications are ineffective. Application of 6pi allows PCSK9 siRNA to efficiently lower plasma cholesterol concentration in vivo, and abolish potentially deleterious off-target phenotypes. The smallest spacer, C3, also shows the same improvement in target specificity. Abasic pivot substitution serves as a general means to harness the specificity of siRNA experiments and therapeutic applications.
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Collections - Graduate School > Department of Life Sciences > 1. Journal Articles
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