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Synthesis and polymerase recognition of a pyrrolocytidine TNA triphosphate
Author(s) -
Mei Hui,
Wang Yajun,
Yik Eric J.,
Chaput John C.
Publication year - 2021
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.23388
Subject(s) - polymerase , nucleic acid , chemistry , dna , dna polymerase , biochemistry , primer (cosmetics) , nuclease , synthetic biology , computational biology , biology , organic chemistry
Synthetic genetics is an area of synthetic biology that aims to extend the properties of heredity and evolution to artificial genetic polymers, commonly known as xeno‐nucleic acids or XNAs. In addition to establishing polymerases that are able to convert genetic information back and forth between DNA and XNA, efforts are underway to construct XNAs with expanded chemical functionality. α‐L‐Threose nucleic acid (TNA), a type of XNA that is recalcitrant to nuclease digestion and amenable to Darwinian evolution, provides a model system for developing XNAs with functional groups that are not present in natural DNA and RNA. Here, we describe the synthesis and polymerase activity of a cytidine TNA triphosphate analog (6‐phenyl‐pyrrolocytosine, tC p TP) that maintains Watson‐Crick base pairing with guanine. Polymerase‐mediated primer extension assays show that tC p TP is an efficient substrate for Kod‐RI, a DNA‐dependent TNA polymerase developed to explore the functional properties of TNA by in vitro selection. Fidelity studies reveal that a cycle of TNA synthesis and reverse transcription occurs with 99.9% overall fidelity when tC p TP and 7‐deaza‐tGTP are present as TNA substrates. This result expands the toolkit of TNA building blocks available for in vitro selection.