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Synthesis and polymerase recognition of a pyrrolocytidine TNA triphosphate.
Biopolymers. 2020 Jul 02 [Online ahead of print]B

Abstract

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, tCp TP) that maintains Watson-Crick base pairing with guanine. Polymerase-mediated primer extension assays show that tCp 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 tCp TP and 7-deaza-tGTP are present as TNA substrates. This result expands the toolkit of TNA building blocks available for in vitro selection.

Authors+Show Affiliations

Departments of Pharmaceutical Sciences, Chemistry, Molecular Biology and Biochemistry, University of California, Irvine, CA, USA. Shenzhen Key Laboratory of Synthetic Genomics, Guangdong Provincial Key Laboratory of Synthetic Genomics, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Science, Shenzhen, China.Departments of Pharmaceutical Sciences, Chemistry, Molecular Biology and Biochemistry, University of California, Irvine, CA, USA.Departments of Pharmaceutical Sciences, Chemistry, Molecular Biology and Biochemistry, University of California, Irvine, CA, USA.Departments of Pharmaceutical Sciences, Chemistry, Molecular Biology and Biochemistry, University of California, Irvine, CA, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32615644

Citation

Mei, Hui, et al. "Synthesis and Polymerase Recognition of a Pyrrolocytidine TNA Triphosphate." Biopolymers, 2020, pp. e23388.
Mei H, Wang Y, Yik EJ, et al. Synthesis and polymerase recognition of a pyrrolocytidine TNA triphosphate. Biopolymers. 2020.
Mei, H., Wang, Y., Yik, E. J., & Chaput, J. C. (2020). Synthesis and polymerase recognition of a pyrrolocytidine TNA triphosphate. Biopolymers, e23388. https://doi.org/10.1002/bip.23388
Mei H, et al. Synthesis and Polymerase Recognition of a Pyrrolocytidine TNA Triphosphate. Biopolymers. 2020 Jul 2;e23388. PubMed PMID: 32615644.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Synthesis and polymerase recognition of a pyrrolocytidine TNA triphosphate. AU - Mei,Hui, AU - Wang,Yajun, AU - Yik,Eric J, AU - Chaput,John C, Y1 - 2020/07/02/ PY - 2020/04/09/received PY - 2020/05/19/revised PY - 2020/05/22/accepted PY - 2020/7/3/entrez PY - 2020/7/3/pubmed PY - 2020/7/3/medline KW - synthetic biology KW - synthetic genetics KW - unnatural nucleobase KW - xeno-nucleic acids SP - e23388 EP - e23388 JF - Biopolymers JO - Biopolymers N2 - 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, tCp TP) that maintains Watson-Crick base pairing with guanine. Polymerase-mediated primer extension assays show that tCp 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 tCp TP and 7-deaza-tGTP are present as TNA substrates. This result expands the toolkit of TNA building blocks available for in vitro selection. SN - 1097-0282 UR - https://www.unboundmedicine.com/medline/citation/32615644/Synthesis_and_polymerase_recognition_of_a_pyrrolocytidine_TNA_triphosphate L2 - https://doi.org/10.1002/bip.23388 DB - PRIME DP - Unbound Medicine ER -
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