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Isoguanine and 5‐Methyl‐Isocytosine Bases, In Vitro and In Vivo
Author(s) -
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
Publication year - 2015
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201406392
Subject(s) - deoxyribose , in vivo , chemistry , nucleic acid , stereochemistry , dna , context (archaeology) , phosphoramidite , deamination , base pair , in vitro , biochemistry , oligonucleotide , biology , enzyme , genetics , paleontology
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.