Fluorescence Probing of DNA Structures in Biology and Nanotechnology
Doktorsavhandling, 2008

Much has been learnt about DNA since its structure was discovered about half a century ago. Scientists everlasting urge to look beyond and deeper into the intricate machinery of life, has continuously pushed technology forward. This Thesis focuses on the development of two new DNA probes, tC and tCO, that both show promises of opening up new possibilities for fluorescence studies of DNA and DNA containing systems. They are fluorescent DNA base analogs, a group of probes with the appealing ability to mimic the natural bases which allows for virtually non‐perturbing substitution into DNA. The cytosine analogs tC and tCO with their minimal effect on the DNA native structure and unique fluorescence properties, offer significant improvements on previously reported base analogs and are especially suited for techniques such as fluorescence resonance energy transfer (FRET), fluorescence anisotropy and fluorescence detected DNA melting (tCO). This Thesis presents a careful characterization of their effect on DNA structure and stability, their photophysical properties, both as monomers and when incorporated into DNA, in addition to demonstrating some examples of their potential as fluorescent DNA probes. The Thesis also explores a new approach to self‐assembled DNA nanoarrays. The approach is intended to allow for the construction of a non‐repetitive and addressable nanoarray with unprecedented information density. As the fundamental pseudohexagonal unit‐cell of this array is significantly smaller than previously published ones, finding ways to characterize this structure has been one of many challenges and a primary focus in this Thesis. The successful and high yield of formation of the fundamental unit was verified using a combination of electrophoresis and spectroscopic techniques. Time‐resolved FRET was shown to be a useful alternative to the microscopy techniques commonly used for larger structures. It was used for retrieving structural information on the pseudo‐hexagon in addition to revealing that this motif has a large conformational flexibility. Details on the formation, melting and stability of the pseudohexagon were obtained by using tCO as a fluorescent probe for DNA melting. This led to a clearer understanding of the thermodynamics governing the formation of the pseudohexagonal structure which will aid in the design and formation of future DNA nanostructures and arrays.

DNA

Fluorescent DNA base analog

UV‐Vis spectroscopy

DNA nanostructure

self‐assembly

time‐resolved fluorescence spectroscopy

KB-salen, Kemigården 4, Chalmers tekniska högskola
Opponent: Prof. David Lilley, University of Dundee, UK

Författare

Peter Sandin

Chalmers, Kemi- och bioteknik, Fysikalisk kemi

Characterization and Use of an Unprecedentedly Bright and Structurally Non-Perturbing Fluorescent DNA Base Analogue

Nucleic Acids Research,; Vol. 36(2008)p. 157-167

Artikel i vetenskaplig tidskrift

Addressable high-information-density DNA nanostructures

Chemical Physics Letters,; Vol. 440(2007)p. 125-129

Artikel i vetenskaplig tidskrift

Synthesis and Oligonucleotide Incorporation of Fluorescent Cytosine Analogue tC: a Promising Nucleic Acid Probe

Nature Protocols,; Vol. 2(2007)p. 615-623

Artikel i vetenskaplig tidskrift

Photophysical Characterization of Fluorescent DNA Base Analogue, tC

Journal of Physical Chemistry B,; Vol. 107(2003)p. 9094-9101

Artikel i vetenskaplig tidskrift

Ämneskategorier

Fysikalisk kemi

ISBN

978-91-7385-099-5

Doktorsavhandlingar vid Chalmers tekniska högskola. Ny serie: 2780

KB-salen, Kemigården 4, Chalmers tekniska högskola

Opponent: Prof. David Lilley, University of Dundee, UK