Cargando…
Isoguanine and 5-Methyl-Isocytosine Bases, In Vitro and In Vivo
The synthesis, base-pairing properties and in vitro and in vivo characteristics of 5-methyl-isocytosine (isoC(Me)) and isoguanine (isoG) nucleosides, incorporated in an HNA(h) (hexitol nucleic acid)–DNA(d) mosaic backbone, are described. The required h-isoG phosphoramidite was prepared by a selectiv...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
WILEY-VCH Verlag
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4531829/ https://www.ncbi.nlm.nih.gov/pubmed/25684598 http://dx.doi.org/10.1002/chem.201406392 |
_version_ | 1782385124238688256 |
---|---|
author | Bande, Omprakash Abu El Asrar, Rania Braddick, Darren Dumbre, Shrinivas Pezo, Valérie Schepers, Guy Pinheiro, Vitor B Lescrinier, Eveline Holliger, Philipp Marlière, Philippe Herdewijn, Piet |
author_facet | Bande, Omprakash Abu El Asrar, Rania Braddick, Darren Dumbre, Shrinivas Pezo, Valérie Schepers, Guy Pinheiro, Vitor B Lescrinier, Eveline Holliger, Philipp Marlière, Philippe Herdewijn, Piet |
author_sort | Bande, Omprakash |
collection | PubMed |
description | The synthesis, base-pairing properties and in vitro and in vivo characteristics of 5-methyl-isocytosine (isoC(Me)) and isoguanine (isoG) nucleosides, incorporated in an HNA(h) (hexitol nucleic acid)–DNA(d) mosaic backbone, are described. The required h-isoG phosphoramidite was prepared by a selective deamination as a key step. As demonstrated by T(m) measurements the hexitol sugar showed slightly better mismatch discrimination against dT. The d-isoG base mispairing follows the order T>G>C while the h-isoG base mispairing follows the order G>C>T. The h- and d-isoC(Me) bases mainly mispair with G. Enzymatic incorporation experiments show that the hexitol backbone has a variable effect on selectivity. In the enzymatic assays, isoG misincorporates mainly with T, and isoC(Me) misincorporates mainly with A. Further analysis in vivo confirmed the patterns of base-pair interpretation for the deoxyribose and hexitol isoC(Me)/isoG bases in a cellular context, through incorporation of the bases into plasmidic DNA. Results in vivo demonstrated that mispairing and misincorporation was dependent on the backbone scaffold of the base, which indicates rational advances towards orthogonality. |
format | Online Article Text |
id | pubmed-4531829 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | WILEY-VCH Verlag |
record_format | MEDLINE/PubMed |
spelling | pubmed-45318292015-08-15 Isoguanine and 5-Methyl-Isocytosine Bases, In Vitro and In Vivo Bande, Omprakash Abu El Asrar, Rania Braddick, Darren Dumbre, Shrinivas Pezo, Valérie Schepers, Guy Pinheiro, Vitor B Lescrinier, Eveline Holliger, Philipp Marlière, Philippe Herdewijn, Piet Chemistry Full Papers The synthesis, base-pairing properties and in vitro and in vivo characteristics of 5-methyl-isocytosine (isoC(Me)) and isoguanine (isoG) nucleosides, incorporated in an HNA(h) (hexitol nucleic acid)–DNA(d) mosaic backbone, are described. The required h-isoG phosphoramidite was prepared by a selective deamination as a key step. As demonstrated by T(m) measurements the hexitol sugar showed slightly better mismatch discrimination against dT. The d-isoG base mispairing follows the order T>G>C while the h-isoG base mispairing follows the order G>C>T. The h- and d-isoC(Me) bases mainly mispair with G. Enzymatic incorporation experiments show that the hexitol backbone has a variable effect on selectivity. In the enzymatic assays, isoG misincorporates mainly with T, and isoC(Me) misincorporates mainly with A. Further analysis in vivo confirmed the patterns of base-pair interpretation for the deoxyribose and hexitol isoC(Me)/isoG bases in a cellular context, through incorporation of the bases into plasmidic DNA. Results in vivo demonstrated that mispairing and misincorporation was dependent on the backbone scaffold of the base, which indicates rational advances towards orthogonality. WILEY-VCH Verlag 2015-03-23 2015-02-13 /pmc/articles/PMC4531829/ /pubmed/25684598 http://dx.doi.org/10.1002/chem.201406392 Text en © 2015 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. https://creativecommons.org/licenses/by/4.0/ © 2015 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Full Papers Bande, Omprakash Abu El Asrar, Rania Braddick, Darren Dumbre, Shrinivas Pezo, Valérie Schepers, Guy Pinheiro, Vitor B Lescrinier, Eveline Holliger, Philipp Marlière, Philippe Herdewijn, Piet Isoguanine and 5-Methyl-Isocytosine Bases, In Vitro and In Vivo |
title | Isoguanine and 5-Methyl-Isocytosine Bases, In Vitro and In Vivo |
title_full | Isoguanine and 5-Methyl-Isocytosine Bases, In Vitro and In Vivo |
title_fullStr | Isoguanine and 5-Methyl-Isocytosine Bases, In Vitro and In Vivo |
title_full_unstemmed | Isoguanine and 5-Methyl-Isocytosine Bases, In Vitro and In Vivo |
title_short | Isoguanine and 5-Methyl-Isocytosine Bases, In Vitro and In Vivo |
title_sort | isoguanine and 5-methyl-isocytosine bases, in vitro and in vivo |
topic | Full Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4531829/ https://www.ncbi.nlm.nih.gov/pubmed/25684598 http://dx.doi.org/10.1002/chem.201406392 |
work_keys_str_mv | AT bandeomprakash isoguanineand5methylisocytosinebasesinvitroandinvivo AT abuelasrarrania isoguanineand5methylisocytosinebasesinvitroandinvivo AT braddickdarren isoguanineand5methylisocytosinebasesinvitroandinvivo AT dumbreshrinivas isoguanineand5methylisocytosinebasesinvitroandinvivo AT pezovalerie isoguanineand5methylisocytosinebasesinvitroandinvivo AT schepersguy isoguanineand5methylisocytosinebasesinvitroandinvivo AT pinheirovitorb isoguanineand5methylisocytosinebasesinvitroandinvivo AT lescriniereveline isoguanineand5methylisocytosinebasesinvitroandinvivo AT holligerphilipp isoguanineand5methylisocytosinebasesinvitroandinvivo AT marlierephilippe isoguanineand5methylisocytosinebasesinvitroandinvivo AT herdewijnpiet isoguanineand5methylisocytosinebasesinvitroandinvivo |