Combined nucleobase and backbone modifications enhance DNA duplex stability and preserve biocompatibility
Combined nucleobase and backbone modifications enhance DNA duplex stability and preserve biocompatibility
DNA strands containing a triazole linkage flanked on its 3?-side by an aminoethylphenoxazine nucleobase analogue (G-clamp) have been prepared by solid-phase synthesis followed by CuAAC-mediated click oligonucleotide ligation. The stability of the doubly modified DNA duplexes and DNA–RNA hybrids is greatly increased, whereas a single base pair mismatch located at or adjacent to the modifications is strongly destabilising, making triazole G-clamp a potent mismatch/point mutation sensor. A DNA strand containing this unnatural combination was successfully amplified by PCR to produce unmodified copies of the original template, with deoxyguanosine inserted opposite to the G-clamp-triazole nucleotide analogue. This study shows for the first time that a polymerase enzyme can read through a combined backbone/nucleobase modification surprisingly well. These favourable properties suggest new applications for oligonucleotides containing the G-clamp triazole modification in biotechnology, nanotechnology, diagnostics and therapeutics.
253-259
El-Sagheer, Afaf H.
05b8295a-64ad-4fdf-ad57-c34934a46c04
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
17 October 2013
El-Sagheer, Afaf H.
05b8295a-64ad-4fdf-ad57-c34934a46c04
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
El-Sagheer, Afaf H. and Brown, Tom
(2013)
Combined nucleobase and backbone modifications enhance DNA duplex stability and preserve biocompatibility.
Chemical Science, 5 (1), .
(doi:10.1039/C3SC51753E).
Abstract
DNA strands containing a triazole linkage flanked on its 3?-side by an aminoethylphenoxazine nucleobase analogue (G-clamp) have been prepared by solid-phase synthesis followed by CuAAC-mediated click oligonucleotide ligation. The stability of the doubly modified DNA duplexes and DNA–RNA hybrids is greatly increased, whereas a single base pair mismatch located at or adjacent to the modifications is strongly destabilising, making triazole G-clamp a potent mismatch/point mutation sensor. A DNA strand containing this unnatural combination was successfully amplified by PCR to produce unmodified copies of the original template, with deoxyguanosine inserted opposite to the G-clamp-triazole nucleotide analogue. This study shows for the first time that a polymerase enzyme can read through a combined backbone/nucleobase modification surprisingly well. These favourable properties suggest new applications for oligonucleotides containing the G-clamp triazole modification in biotechnology, nanotechnology, diagnostics and therapeutics.
This record has no associated files available for download.
More information
Published date: 17 October 2013
Additional Information:
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence.
Organisations:
Chemistry
Identifiers
Local EPrints ID: 369278
URI: http://eprints.soton.ac.uk/id/eprint/369278
ISSN: 1478-6524
PURE UUID: 64c9d75b-a035-4664-a3c7-b2d975650c5c
Catalogue record
Date deposited: 23 Sep 2014 10:32
Last modified: 15 Mar 2024 03:25
Export record
Altmetrics
Contributors
Author:
Afaf H. El-Sagheer
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics