Fluorescent DNA nanotags based on a self-assembled DNA tetrahedron.

Progress in fluorescence detection and imaging technologies depends on the availability of fluorescent labels with strong light absorption/emission characteristics. We have synthesized intercalator dye arrays on a compact 3-dimensional DNA-tetrahedron nanostructure. The template tolerates the structural distortions introduced by intercalation and allows concentration of multiple fluorophores within a small volume, resulting in brightly fluorescent nanotags with effective extinction coefficients in the order of 10(6) M(-1) cm(-1). Efficient energy transfer from intercalated donor dyes to covalently attached acceptor dyes in the nanotags allows the emission wavelength to be shifted to the red relative to the excitation light, providing wavelength tunability. The compact nature of the supramolecular DNA tetrahedron also provides a protective medium for the fluorophores, leading to improved photostability and enhanced resistance to nuclease digestion, relative to one- or two-dimensional nanotags described previously.

[1]  L. Lerman,et al.  Structural considerations in the interaction of DNA and acridines. , 1961, Journal of molecular biology.

[2]  N. Seeman DNA nanotechnology: novel DNA constructions. , 1998, Annual review of biophysics and biomolecular structure.

[3]  Andrew J Turberfield,et al.  The single-step synthesis of a DNA tetrahedron. , 2004, Chemical communications.

[4]  Weihong Tan,et al.  Multicolor FRET silica nanoparticles by single wavelength excitation. , 2006, Nano letters.

[5]  James J. Russo,et al.  Combinatorial fluorescence energy transfer tags for multiplex biological assays , 2001, Nature Biotechnology.

[6]  K. Hahn,et al.  Solvent-sensitive dyes to report protein conformational changes in living cells. , 2003, Journal of the American Chemical Society.

[7]  Thomas Carell,et al.  Click chemistry as a reliable method for the high-density postsynthetic functionalization of alkyne-modified DNA. , 2006, Organic letters.

[8]  D. Crothers Calculation of binding isotherms for heterogeneous polymers , 1968, Biopolymers.

[9]  T. Netzel,et al.  Base-Content Dependence of Emission Enhancements, Quantum Yields, and Lifetimes for Cyanine Dyes Bound to Double-Strand DNA: Photophysical Properties of Monomeric and Bichromomphoric DNA Stains , 1995 .

[10]  B. Roques,et al.  DNA Bifunctional intercalators. 2. Fluorescence properties and DNA binding interaction of an ethidium homodimer and an acridine ethidium heterodimer. , 1978, Biochemistry.

[11]  D. Schwartz,et al.  Inhibition of restriction endonuclease activity by DNA binding fluorochromes. , 1996, Journal of biomolecular structure & dynamics.

[12]  Nadrian C Seeman,et al.  At the crossroads of chemistry, biology, and materials: structural DNA nanotechnology. , 2003, Chemistry & biology.

[13]  Mats Jonsson,et al.  Characterization of the Binding of the Fluorescent Dyes Yo and Yoyo to DNA by Polarized-Light Spectroscopy , 1994 .

[14]  P. Sima,et al.  Structural and Environmental Requirements for Quenching of Singlet Oxygen by Cyanine Dyes† , 2000, Photochemistry and photobiology.

[15]  D E Wemmer,et al.  Stable fluorescent complexes of double-stranded DNA with bis-intercalating asymmetric cyanine dyes: properties and applications. , 1992, Nucleic acids research.

[16]  Multiple labeling of antibodies with dye/DNA conjugate for sensitivity improvement in fluorescence immunoassay. , 2007, Bioconjugate chemistry.

[17]  Christopher L. Brown,et al.  Rapid purification of EGFP, EYFP, and ECFP with high yield and purity. , 2005, Protein expression and purification.

[18]  A. Glazer,et al.  Light guides. Directional energy transfer in a photosynthetic antenna. , 1989, The Journal of biological chemistry.

[19]  William M. Shih,et al.  A 1.7-kilobase single-stranded DNA that folds into a nanoscale octahedron , 2004, Nature.

[20]  V. Balzani,et al.  Light-harvesting dendrimers. , 2003, Current opinion in chemical biology.

[21]  P. Rothemund Folding DNA to create nanoscale shapes and patterns , 2006, Nature.

[22]  B. Åkerman,et al.  Photobleaching of asymmetric cyanines used for fluorescence imaging of single DNA molecules. , 2001, Journal of the American Chemical Society.

[23]  E. Vauthey,et al.  Ultrafast excited-state dynamics of DNA fluorescent intercalators: new insight into the fluorescence enhancement mechanism. , 2006, Journal of the American Chemical Society.

[24]  J. Petty,et al.  Thermodynamic Characterization of the Association of Cyanine Dyes with DNA , 2000 .

[25]  Alan Waggoner,et al.  Fluorescent labels for proteomics and genomics. , 2006, Current opinion in chemical biology.

[26]  R. Haugland,et al.  Green- and red-fluorescent nanospheres for the detection of cell surface receptors by flow cytometry. , 1998, Journal of immunological methods.

[27]  Elizabeth A Jares-Erijman,et al.  Imaging molecular interactions in living cells by FRET microscopy. , 2006, Current opinion in chemical biology.

[28]  A. Waggoner,et al.  Fluorescent DNA nanotags: supramolecular fluorescent labels based on intercalating dye arrays assembled on nanostructured DNA templates. , 2007, Journal of the American Chemical Society.

[29]  S. Quake,et al.  Nanometer-scale Fluorescence Resonance Optical Waveguides , 2004 .

[30]  Russell P. Goodman,et al.  Rapid Chiral Assembly of Rigid DNA Building Blocks for Molecular Nanofabrication , 2005, Science.

[31]  J. Jacobsen,et al.  1H NMR studies of the bis-intercalation of a homodimeric oxazole yellow dye in DNA oligonucleotides. , 1998, Journal of biomolecular structure & dynamics.

[32]  Dan Luo,et al.  Multiplexed detection of pathogen DNA with DNA-based fluorescence nanobarcodes , 2005, Nature Biotechnology.

[33]  Nadrian C Seeman,et al.  Assembly and characterization of 8-arm and 12-arm DNA branched junctions. , 2007, Journal of the American Chemical Society.

[34]  Boris Rybtchinski,et al.  Combining light-harvesting and charge separation in a self-assembled artificial photosynthetic system based on perylenediimide chromophores. , 2004, Journal of the American Chemical Society.

[35]  C. Niemeyer REVIEW Nanoparticles, Proteins, and Nucleic Acids: Biotechnology Meets Materials Science , 2022 .

[36]  Joby Eldo,et al.  Improved photostability and fluorescence properties through polyfluorination of a cyanine dye. , 2004, Organic letters.