Assembling of G-strands into novel tetra-molecular parallel G4-DNA nanostructures using avidin-biotin recognition

Nucleic Acids Res. 2008 Sep;36(15):5050-60. doi: 10.1093/nar/gkn459. Epub 2008 Jul 28.

Abstract

We describe a method for the preparation of novel long (hundreds of nanometers), uniform, inter-molecular G4-DNA molecules composed of four parallel G-strands. The only long continuous G4-DNA reported so far are intra-molecular structures made of a single G-strand. To enable a tetra-molecular assembly of the G-strands we developed a novel approach based on avidin-biotin biological recognition. The steps of the G4-DNA production include: (i) Enzymatic synthesis of long poly(dG)-poly(dC) molecules with biotinylated poly(dG)-strand; (ii) Formation of a complex between avidin-tetramer and four biotinylated poly(dG)-poly(dC) molecules; (iii) Separation of the poly(dC) strands from the poly(dG)-strands, which are connected to the avidin; (iv) Assembly of the four G-strands attached to the avidin into tetra-molecular G4-DNA. The average contour length of the formed structures, as measured by AFM, is equal to that of the initial poly(dG)-poly(dC) molecules, suggesting a tetra-molecular mechanism of the G-strands assembly. The height of tetra-molecular G4-nanostructures is larger than that of mono-molecular G4-DNA molecules having similar contour length. The CD spectra of the tetra- and mono-molecular G4-DNA are markedly different, suggesting different structural organization of these two types of molecules. The tetra-molecular G4-DNA nanostructures showed clear electrical polarizability. This suggests that they may be useful for molecular electronics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Avidin / chemistry*
  • Avidin / ultrastructure
  • Biotin / chemistry*
  • Circular Dichroism
  • G-Quadruplexes*
  • Microscopy, Atomic Force
  • Nanostructures / chemistry*
  • Nanostructures / ultrastructure
  • Poly C / chemistry
  • Poly G / chemistry
  • Polydeoxyribonucleotides / chemistry*
  • Static Electricity

Substances

  • Polydeoxyribonucleotides
  • Avidin
  • Poly G
  • poly(dC)
  • poly(dG)
  • Poly C
  • Biotin