Cotranscriptional Production of Chemically Modified RNA Nanoparticles

Methods Mol Biol. 2017;1632:91-105. doi: 10.1007/978-1-4939-7138-1_6.

Abstract

RNA nanoparticles consisting of multiple RNA strands of different sequences forming various three-dimensional structures emerge as promising carriers of siRNAs, RNA aptamers, and ribozymes. In vitro transcription of a mixture of dsDNA templates encoding all the subunits of the RNA nanoparticle may result in cotranscriptional self-assembly of the nanoparticle. Based on our experience with production of RNA nanorings, RNA nanocubes, and RNA three-way junctions, we propose a strategy for optimization of the cotranscriptional production of chemically modified ribonuclease-resistant RNA nanoparticles.

Keywords: 2′-F-dCTP; 2′-F-dUTP; Cotranscriptional assembly; In vitro transcription; RNA nanocubes; RNA nanorings; T7 R&DNA polymerase; T7 RNA polymerase.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • 5' Untranslated Regions
  • Base Sequence
  • Halogenation
  • Mutation
  • Nanoparticles*
  • RNA / chemistry*
  • RNA / genetics*
  • Transcription, Genetic*

Substances

  • 5' Untranslated Regions
  • RNA