Folding RNA-Protein Complex into Designed Nanostructures

Methods Mol Biol. 2015;1316:169-79. doi: 10.1007/978-1-4939-2730-2_14.

Abstract

RNA-protein (RNP) complexes are promising biomaterials for the fields of nanotechnology and synthetic biology. Protein-responsive RNA sequences (RNP motifs) can be integrated into various RNAs, such as messenger RNA, short-hairpin RNA, and synthetic RNA nano-objects for a variety of purposes. Direct observation of RNP interaction in solution at high resolution is important in the design and construction of RNP-mediated nanostructures. Here we describe a method to construct and visualize RNP nanostructures that precisely arrange a target protein on the RNA scaffold with nanometer scale. High-speed AFM (HS-AFM) images of RNP nanostructures show that the folding of RNP complexes of defined sizes can be directly visualized at single RNP resolution in solution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electrophoretic Mobility Shift Assay / methods
  • Microscopy, Atomic Force
  • Nanostructures / chemistry*
  • Nucleic Acid Conformation*
  • RNA / chemical synthesis
  • RNA / chemistry*
  • RNA / metabolism
  • Ribonucleoproteins / chemical synthesis
  • Ribonucleoproteins / chemistry*
  • Ribonucleoproteins / metabolism

Substances

  • Ribonucleoproteins
  • RNA