Enzymatic Synthesis of Self-assembled Dicer Substrate RNA Nanostructures for Programmable Gene Silencing

Bora Jang, Boyoung Kim, Hyunsook Kim, Hyokyoung Kwon, Minjeong Kim, Yunmi Seo, Marion Colas, Hansaem Jeong, Eun Hye Jeong, Kyuri Lee, Hyukjin Lee

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

Enzymatic synthesis of RNA nanostructures is achieved by isothermal rolling circle transcription (RCT). Each arm of RNA nanostructures provides a functional role of Dicer substrate RNA inducing sequence specific RNA interference (RNAi). Three different RNAi sequences (GFP, RFP, and BFP) are incorporated within the three-arm junction RNA nanostructures (Y-RNA). The template and helper DNA strands are designed for the large-scale in vitro synthesis of RNA strands to prepare self-assembled Y-RNA. Interestingly, Dicer processing of Y-RNA is highly influenced by its physical structure and different gene silencing activity is achieved depending on its arm length and overhang. In addition, enzymatic synthesis allows the preparation of various Y-RNA structures using a single DNA template offering on demand regulation of multiple target genes.

Original languageEnglish
Pages (from-to)4279-4284
Number of pages6
JournalNano Letters
Volume18
Issue number7
DOIs
StatePublished - 11 Jul 2018

Keywords

  • Dicer substrate RNA
  • enzymatic synthesis
  • gene silencing
  • programmable gene regulation
  • RNA nanostructures

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