Abasic pivot substitution harnesses target specificity of RNA interference

Hye Sook Lee, Heeyoung Seok, Dong Ha Lee, Juyoung Ham, Wooje Lee, Emilia Moonkyung Youm, Jin Seon Yoo, Yong Seung Lee, Eun Sook Jang, Sung Wook Chi

Research output: Contribution to journalArticle

15 Citations (Scopus)

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 ∼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.

Original languageEnglish
Article number10154
JournalNature communications
Volume6
DOIs
Publication statusPublished - 2015 Dec 18

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

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    Lee, H. S., Seok, H., Lee, D. H., Ham, J., Lee, W., Youm, E. M., Yoo, J. S., Lee, Y. S., Jang, E. S., & Chi, S. W. (2015). Abasic pivot substitution harnesses target specificity of RNA interference. Nature communications, 6, [10154]. https://doi.org/10.1038/ncomms10154