An in vitro selection protocol for threose nucleic acid (TNA) using DNA display

Matthew R. Dunn, John C. Chaput

Research output: Contribution to journalArticlepeer-review

6 Scopus citations


Threose nucleic acid (TNA) is an unnatural genetic polymer composed of repeating threofuranosyl sugars linked by 2' and 3' phosphodiester bonds. TNA is capable of forming antiparallel Watson-Crick duplex structures in a self-pairing mode, and can also cross-pair opposite complementary strands of DNA and RNA. The solution NMR structure of a self-complementary TNA duplex reveals that TNA adopts an A-form helical structure, which explains its ability to exchange genetic information with natural genetic polymers. In a recent advance, a TNA aptamer was evolved from a pool of random sequences using an engineered polymerase that can copy DNA sequences into TNA. This unit details the steps required to evolve functional TNA molecules in the laboratory using a method called DNA display. Using this approach, TNA molecules are physically linked to their encoding double-stranded DNA template. By linking TNA phenotype with DNA genotype, one can select for TNA molecules with a desired function and recover their encoding genetic information by PCR amplification. Each round of selection requires ~3 days to complete and multiple rounds of selection and amplification are required to generate functional TNA molecules.

Original languageEnglish (US)
Pages (from-to)9.8.1-9.8.19
JournalCurrent Protocols in Nucleic Acid Chemistry
StatePublished - 2014


  • Aptamer
  • DNA display
  • In vitro selection
  • Oligonucleotide
  • Threose nucleic acid (TNA)

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry


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