Bernardes Pinheiro, VB;
(2018)
Phosphonomethyl Oligonucleotides as Backbone Modified Artificial Genetic Polymers.
Journal of the American Chemical Society
10.1021/jacs.8b03447.
(In press).
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Abstract
Although several synthetic or xenobiotic nucleic acids (XNAs) have been shown to be viable genetic materials in vitro, major hurdles remain for their in vivo applications, particularly orthogonality. The availability of XNAs that do not interact with natural nucleic acids and are not affected by natural DNA processing enzymes, as well as specialized XNA processing enzymes that do not interact with natural nucleic acids, are essential. Here, we report 3’–2’ phosphonomethyl-threosyl nu-cleic acid (tPhoNA) as a novel XNA genetic material and a prime candidate for in vivo XNA applications. We established routes for the chemical synthesis of phosphonate nucleic acids and phosphorylated monomeric building blocks, and we demonstrated that DNA duplexes were destabilized upon replacement with tPhoNA. We engineered a novel tPhoNA synthe-tase enzyme and, with a previously reported XNA reverse transcriptase, demonstrated that tPhoNA is a viable genetic mate-rial (with an aggregate error rate of approximately 17 x 10-3 per base) compatible with the isolation of functional XNAs. In vivo experiments to test tPhoNA orthogonality showed that the E. coli cellular machinery had only very limited potential to access genetic information in tPhoNA. Our work is the first report of a synthetic genetic material modified in both sugar and phosphate backbone moieties and represents a significant advance in biorthogonality towards the introduction of XNA systems in vivo.
Type: | Article |
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Title: | Phosphonomethyl Oligonucleotides as Backbone Modified Artificial Genetic Polymers |
Open access status: | An open access version is available from UCL Discovery |
DOI: | 10.1021/jacs.8b03447 |
Publisher version: | http://doi.org/10.1021/jacs.8b03447 |
Language: | English |
Additional information: | © 2018 American Chemical Society. This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
Keywords: | XNA; Phosphono nucleic acid; Polymerase evolution; In vivo transliteration; H-Phosphinates; Xenobiology |
UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology |
URI: | https://discovery.ucl.ac.uk/id/eprint/10047973 |
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