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Characterization and use of an unprecedentedly bright and structurally non-perturbing fluorescent DNA base analogue

Characterization and use of an unprecedentedly bright and structurally non-perturbing fluorescent DNA base analogue
Characterization and use of an unprecedentedly bright and structurally non-perturbing fluorescent DNA base analogue
This article presents the first evidence that the DNA base analogue 1,3-diaza-2-oxophenoxazine, tCO, is highly fluorescent, both as free nucleoside and incorporated in an arbitrary DNA structure. tCO is thoroughly characterized with respect to its photophysical properties and structural performance in single- and double-stranded oligonucleotides. The lowest energy absorption band at 360 nm ({varepsilon} = 9000 M–1 cm–1) is dominated by a single in-plane polarized electronic transition and the fluorescence, centred at 465 nm, has a quantum yield of 0.3. When incorporated into double-stranded DNA, tCO shows only minor variations in fluorescence intensity and lifetime with neighbouring bases, and the average quantum yield is 0.22. These features make tCO, on average, the brightest DNA-incorporated base analogue so far reported. Furthermore, it base pairs exclusively with guanine and causes minimal perturbations to the native structure of DNA. These properties make tCO a promising base analogue that is perfectly suited for e.g. photophysical studies of DNA interacting with macromolecules (proteins) or for determining size and shape of DNA tertiary structures using techniques such as fluorescence anisotropy and fluorescence resonance energy transfer (FRET)
0305-1048
157-167
Sandin, Peter
f7324993-b21e-4575-ae67-fd3223624030
Borjesson, Karl
f2524b10-bda3-4a92-b799-920a641135ca
Li, Hong
44d51d43-1fe5-4d15-820d-22d302b9e1a7
Martensson, Jerker
2623c3f1-c729-4d58-8488-64a0b1aa68f7
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Wilhelmsson, L. Marcus
bcf4f67a-198c-4c5e-80b8-a78f743509d4
Albinsson, Bo
1571067f-df46-43f7-96b9-52dc43278777
Sandin, Peter
f7324993-b21e-4575-ae67-fd3223624030
Borjesson, Karl
f2524b10-bda3-4a92-b799-920a641135ca
Li, Hong
44d51d43-1fe5-4d15-820d-22d302b9e1a7
Martensson, Jerker
2623c3f1-c729-4d58-8488-64a0b1aa68f7
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Wilhelmsson, L. Marcus
bcf4f67a-198c-4c5e-80b8-a78f743509d4
Albinsson, Bo
1571067f-df46-43f7-96b9-52dc43278777

Sandin, Peter, Borjesson, Karl, Li, Hong, Martensson, Jerker, Brown, Tom, Wilhelmsson, L. Marcus and Albinsson, Bo (2008) Characterization and use of an unprecedentedly bright and structurally non-perturbing fluorescent DNA base analogue. Nucleic Acids Research, 36 (1), 157-167. (doi:10.1093/nar/gkm1006). (PMID:18003656)

Record type: Article

Abstract

This article presents the first evidence that the DNA base analogue 1,3-diaza-2-oxophenoxazine, tCO, is highly fluorescent, both as free nucleoside and incorporated in an arbitrary DNA structure. tCO is thoroughly characterized with respect to its photophysical properties and structural performance in single- and double-stranded oligonucleotides. The lowest energy absorption band at 360 nm ({varepsilon} = 9000 M–1 cm–1) is dominated by a single in-plane polarized electronic transition and the fluorescence, centred at 465 nm, has a quantum yield of 0.3. When incorporated into double-stranded DNA, tCO shows only minor variations in fluorescence intensity and lifetime with neighbouring bases, and the average quantum yield is 0.22. These features make tCO, on average, the brightest DNA-incorporated base analogue so far reported. Furthermore, it base pairs exclusively with guanine and causes minimal perturbations to the native structure of DNA. These properties make tCO a promising base analogue that is perfectly suited for e.g. photophysical studies of DNA interacting with macromolecules (proteins) or for determining size and shape of DNA tertiary structures using techniques such as fluorescence anisotropy and fluorescence resonance energy transfer (FRET)

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Published date: January 2008

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Local EPrints ID: 146713
URI: http://eprints.soton.ac.uk/id/eprint/146713
ISSN: 0305-1048
PURE UUID: 1f1b1687-7a0d-4fa2-be36-66610f8e82a9

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Date deposited: 09 Jun 2010 08:08
Last modified: 17 Jul 2019 00:04

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