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An efficient and faithful in vitro replication system for threose nucleic acid.
J Am Chem Soc 2013; 135(9):3583-91JA

Abstract

The emerging field of synthetic genetics provides an opportunity to explore the structural and functional properties of synthetic genetic polymers by in vitro selection. Limiting this process, however, is the availability of enzymes that allow for the synthesis and propagation of genetic information present in unnatural nucleic acid sequences. Here, we report the development of a transcription and reverse-transcription system that can replicate unnatural genetic polymers composed of threose nucleic acids (TNA). TNA is a potential progenitor of RNA in which the natural ribose sugar found in RNA has been replaced with an unnatural threose sugar. Using commercial polymerases that recognize TNA, we demonstrate that an unbiased three-letter and two different biased four-letter genetic alphabets replicate in vitro with high efficiency and high overall fidelity. We validated the replication system by performing one cycle of transcription, selection, reverse transcription, and amplification on a library of 10(14) DNA templates and observed ∼380-fold enrichment after one round of selection for a biotinylated template. We further show that TNA polymers are stable to enzymes that degrade DNA and RNA. These results provide the methodology needed to evolve biologically stable aptamers and enzymes for exobiology and molecular medicine.

Authors+Show Affiliations

Center for Evolutionary Medicine and Informatics in the Biodesign Institute and Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287-5301, United States.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23432469

Citation

Yu, Hanyang, et al. "An Efficient and Faithful in Vitro Replication System for Threose Nucleic Acid." Journal of the American Chemical Society, vol. 135, no. 9, 2013, pp. 3583-91.
Yu H, Zhang S, Dunn MR, et al. An efficient and faithful in vitro replication system for threose nucleic acid. J Am Chem Soc. 2013;135(9):3583-91.
Yu, H., Zhang, S., Dunn, M. R., & Chaput, J. C. (2013). An efficient and faithful in vitro replication system for threose nucleic acid. Journal of the American Chemical Society, 135(9), pp. 3583-91. doi:10.1021/ja3118703.
Yu H, et al. An Efficient and Faithful in Vitro Replication System for Threose Nucleic Acid. J Am Chem Soc. 2013 Mar 6;135(9):3583-91. PubMed PMID: 23432469.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An efficient and faithful in vitro replication system for threose nucleic acid. AU - Yu,Hanyang, AU - Zhang,Su, AU - Dunn,Matthew R, AU - Chaput,John C, Y1 - 2013/02/25/ PY - 2013/2/26/entrez PY - 2013/2/26/pubmed PY - 2013/9/4/medline SP - 3583 EP - 91 JF - Journal of the American Chemical Society JO - J. Am. Chem. Soc. VL - 135 IS - 9 N2 - The emerging field of synthetic genetics provides an opportunity to explore the structural and functional properties of synthetic genetic polymers by in vitro selection. Limiting this process, however, is the availability of enzymes that allow for the synthesis and propagation of genetic information present in unnatural nucleic acid sequences. Here, we report the development of a transcription and reverse-transcription system that can replicate unnatural genetic polymers composed of threose nucleic acids (TNA). TNA is a potential progenitor of RNA in which the natural ribose sugar found in RNA has been replaced with an unnatural threose sugar. Using commercial polymerases that recognize TNA, we demonstrate that an unbiased three-letter and two different biased four-letter genetic alphabets replicate in vitro with high efficiency and high overall fidelity. We validated the replication system by performing one cycle of transcription, selection, reverse transcription, and amplification on a library of 10(14) DNA templates and observed ∼380-fold enrichment after one round of selection for a biotinylated template. We further show that TNA polymers are stable to enzymes that degrade DNA and RNA. These results provide the methodology needed to evolve biologically stable aptamers and enzymes for exobiology and molecular medicine. SN - 1520-5126 UR - https://www.unboundmedicine.com/medline/citation/23432469/An_efficient_and_faithful_in_vitro_replication_system_for_threose_nucleic_acid_ L2 - https://dx.doi.org/10.1021/ja3118703 DB - PRIME DP - Unbound Medicine ER -