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

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 assemb...

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Bibliographic Details
Published in:Nucleic acids research 2008-09, Vol.36 (15), p.5050-5060
Main Authors: Borovok, Natalia, Iram, Natalie, Zikich, Dragoslav, Ghabboun, Jamal, Livshits, Gideon I, Porath, Danny, Kotlyar, Alexander B
Format: Article
Language:English
Subjects:
Avidin - chemistry
Avidin - ultrastructure
Biotin - chemistry
Chemistry and Synthetic Biology
Circular Dichroism
G-Quadruplexes
Microscopy, Atomic Force
Nanostructures - chemistry
Nanostructures - ultrastructure
Poly C - chemistry
Poly G - chemistry
Polydeoxyribonucleotides - chemistry
Static Electricity
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Summary: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.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gkn459