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RNA Scaffolds pp 169-179 | Cite as

Folding RNA–Protein Complex into Designed Nanostructures

  • Tomonori Shibata
  • Yuki Suzuki
  • Hiroshi Sugiyama
  • Masayuki Endo
  • Hirohide SaitoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1316)

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.

Key words

Ribonucleoprotein Kink-turn RNA L7Ae RNA–protein complex RNA nanostructures RNA nanotechnology RNP High-speed atomic force microscopy 

Notes

Acknowledgement

We thank Hirohisa Ohno and Eriko Osada (Kyoto University) to initiate the projects. We also thank Peter Karagiannis (Kyoto University) for critically reading the manuscript. This work was supported by the New Energy and Industrial Technology Development Organization (09A02021a) and a Grant-in-Aid for Scientific Research on Innovative Areas “Molecular Robotics” (No. 24104002) from The Ministry of Education, Culture, Sport, Science, and Technology, Japan.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Tomonori Shibata
    • 2
  • Yuki Suzuki
    • 1
  • Hiroshi Sugiyama
    • 1
  • Masayuki Endo
    • 3
  • Hirohide Saito
    • 2
    Email author
  1. 1.Department of Chemistry, Graduate School of ScienceKyoto UniversityKyotoJapan
  2. 2.Center for iPS Cell Research and Application (CiRA)Kyoto UniversityKyotoJapan
  3. 3.Institute for Integrated Cell-Material Sciences (WPI-iCeMS)Kyoto UniversityKyotoJapan

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