Fabrication of stable and RNase-resistant RNA nanoparticles active in gearing the nanomotors for viral DNA packaging

ACS Nano. 2011 Jan 25;5(1):237-46. doi: 10.1021/nn1024658. Epub 2010 Dec 14.

Abstract

Both DNA and RNA can serve as powerful building blocks for bottom-up fabrication of nanostructures. A pioneering concept proposed by Ned Seeman 30 years ago has led to an explosion of knowledge in DNA nanotechnology. RNA can be manipulated with simplicity characteristic of DNA, while possessing noncanonical base-pairing, versatile function, and catalytic activity similar to proteins. However, standing in awe of the sensitivity of RNA to RNase degradation has made many scientists flinch away from RNA nanotechnology. Here we report the construction of stable RNA nanoparticles resistant to RNase digestion. The 2'-F (2'-fluoro) RNA retained its property for correct folding in dimer formation, appropriate structure in procapsid binding, and biological activity in gearing the phi29 nanomotor to package viral DNA and producing infectious viral particles. Our results demonstrate that it is practical to produce RNase-resistant, biologically active, and stable RNA for application in nanotechnology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacillus Phages / genetics
  • Bacillus Phages / metabolism
  • Bacillus Phages / physiology
  • Base Sequence
  • Cattle
  • DNA Packaging*
  • DNA, Viral / genetics*
  • DNA-Directed RNA Polymerases / genetics
  • DNA-Directed RNA Polymerases / metabolism
  • Dimerization
  • Hydrogen-Ion Concentration
  • Models, Molecular
  • Mutation
  • Nanoparticles / chemistry*
  • Nanotechnology / methods*
  • Nucleic Acid Conformation
  • Pyrimidines / chemistry
  • RNA / chemistry*
  • RNA / genetics
  • RNA / isolation & purification
  • RNA / metabolism
  • RNA Stability*
  • Ribonucleases / metabolism*
  • Transcription, Genetic
  • Viral Proteins / genetics
  • Viral Proteins / metabolism
  • Virion / metabolism
  • Virus Assembly

Substances

  • DNA, Viral
  • Pyrimidines
  • Viral Proteins
  • RNA
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases
  • Ribonucleases
  • pyrimidine