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Enzymatic Construction of Artificial Base Pairs: The Effect of Metal Shielding

Abstract : Formation of metal base pairs is a versatile method for the introduction of metal cations into nucleic acids that has been applied in numerous applications including the construction of metal nanowires, development of energy, charge transfer devices and expansion of the genetic alphabet. As an alternative, enzymatic construction of metal base pairs represents an alluring strategy that grants access to longer sequences and offers the possibility of using such unnatural base pairs (UBPs) in SELEX experiments for the identification of functional nucleic acids. This method remains rather underexplored and a better understanding of the key parameters in the design of efficient nucleotides is required. Herein, we have investigated the effect of methylation of the imidazole nucleoside (dIm nMe TP) on the efficiency of the enzymatic construction of metal base pairs. The presence of methyl substituents on dImTP facilitates the polymerase-driven formation of dIm 4Me-Ag I-dIm and dIm 2Me TP-Cr III-dIm base pairs. Steric factors rather than basicity of the imidazole nucleobase appear to govern the enzymatic formation of such metal base pairs. We also demonstrate the compatibility of other metal cations rarely considered in the construction of artificial metal base with enzymatic DNA synthesis under both primer extension reaction and PCR conditions. These findings open up new directions for the design of nucleotide analogs for the development of metal base pairs. 2
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Submitted on : Wednesday, September 2, 2020 - 2:32:02 PM
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Marie Flamme, Fabienne Levi-Acobas, Susanne Hensel, Shuvankar Naskar, Pascal Röthlisberger, et al.. Enzymatic Construction of Artificial Base Pairs: The Effect of Metal Shielding. ChemBioChem, 2020, ⟨10.1002/cbic.202000402⟩. ⟨pasteur-02928354⟩



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