Base-Modified Nucleic Acids as a Powerful Tool for Synthetic Biology and Biotechnology

Chemistry : a European Journal
Elena EremeevaPiet Herdewijn

Abstract

The ability of various nucleoside triphosphate analogues of deoxyguanosine and deoxycytidine with 7-deazadeoxyadenosine (A1 ) and 5-chlorodeoxyuridine (T1 ) to serve as substrates for Taq DNA polymerase was evaluated. The triphosphate set composed of A1 , T1 , and 7-deazadeoxyguanosine with either 5-methyldeoxycytidine or 5-fluorodeoxycytidine was successfully employed in the polymerase chain reaction (PCR) of 1.5 kb fragments as well as random oligonucleotide libraries. Another effective combination of triphosphates for the synthesis of a 1 kb PCR product was A1 , T1 , deoxyinosine, and 5-bromodeoxycytidine. In vivo experiments using an antibiotic-resistant gene containing the latter set demonstrated that the bacterial machinery accepts fully modified sequences as genetic templates. Moreover, the ability of the base-modified segments to selectively protect DNA from cleavage by restriction endonucleases was shown. This approach can be used to regulate the endonuclease cleavage pattern.

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Citations

Nov 17, 2017·International Journal of Molecular Sciences·Pascal RöthlisbergerMarcel Hollenstein
Dec 24, 2019·Chembiochem : a European Journal of Chemical Biology·Hoai NguyenPiet Herdewijn
Jul 25, 2018·Current Protocols in Chemical Biology·Elena Eremeeva, Piet Herdewijn
Aug 6, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Peng NieHui Mei
Dec 11, 2020·Nucleic Acids Research·Asem AlenaizanAnton S Petrov
May 1, 2021·RSC Chemical Biology·Adeline EspinasseErin E Carlson

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