An efficient and faithful in vitro replication system for threose nucleic acid

Hanyang Yu, Su Zhang, Matthew R. Dunn, John C. Chaput

Research output: Contribution to journalArticle

44 Citations (Scopus)

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 1014 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.

Original languageEnglish (US)
Pages (from-to)3583-3591
Number of pages9
JournalJournal of the American Chemical Society
Volume135
Issue number9
DOIs
StatePublished - Mar 6 2013

Fingerprint

Nucleic acids
Transcription
Nucleic Acids
RNA
Enzymes
Polymers
Sugars
DNA
Astrobiology
Nucleic acid sequences
Reverse Transcription
Exobiology
Molecular Medicine
Ribose
Medicine
Amplification
Gene Library
Availability
erythrose
In Vitro Techniques

ASJC Scopus subject areas

  • Chemistry(all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

An efficient and faithful in vitro replication system for threose nucleic acid. / Yu, Hanyang; Zhang, Su; Dunn, Matthew R.; Chaput, John C.

In: Journal of the American Chemical Society, Vol. 135, No. 9, 06.03.2013, p. 3583-3591.

Research output: Contribution to journalArticle

Yu, Hanyang ; Zhang, Su ; Dunn, Matthew R. ; Chaput, John C. / An efficient and faithful in vitro replication system for threose nucleic acid. In: Journal of the American Chemical Society. 2013 ; Vol. 135, No. 9. pp. 3583-3591.
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