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An In Vitro Selection Protocol for Threose Nucleic Acid (TNA) Using DNA Display.
Curr Protoc Nucleic Acid Chem 2014; 57:9.8.1-19CP

Abstract

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.

Authors+Show Affiliations

School of Life Sciences, Arizona State University, Tempe, Arizona; The Biodesign Institute at Arizona State University, Tempe, Arizona.No affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

24961723

Citation

Dunn, Matthew R., and John C. Chaput. "An in Vitro Selection Protocol for Threose Nucleic Acid (TNA) Using DNA Display." Current Protocols in Nucleic Acid Chemistry, vol. 57, 2014, pp. 9.8.1-19.
Dunn MR, Chaput JC. An In Vitro Selection Protocol for Threose Nucleic Acid (TNA) Using DNA Display. Curr Protoc Nucleic Acid Chem. 2014;57:9.8.1-19.
Dunn, M. R., & Chaput, J. C. (2014). An In Vitro Selection Protocol for Threose Nucleic Acid (TNA) Using DNA Display. Current Protocols in Nucleic Acid Chemistry, 57, pp. 9.8.1-19. doi:10.1002/0471142700.nc0908s57.
Dunn MR, Chaput JC. An in Vitro Selection Protocol for Threose Nucleic Acid (TNA) Using DNA Display. Curr Protoc Nucleic Acid Chem. 2014 Jun 24;57:9.8.1-19. PubMed PMID: 24961723.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An In Vitro Selection Protocol for Threose Nucleic Acid (TNA) Using DNA Display. AU - Dunn,Matthew R, AU - Chaput,John C, Y1 - 2014/06/24/ PY - 2014/6/26/entrez PY - 2014/6/26/pubmed PY - 2016/6/28/medline KW - DNA display KW - aptamer KW - in vitro selection KW - oligonucleotide KW - threose nucleic acid (TNA) SP - 9.8.1 EP - 19 JF - Current protocols in nucleic acid chemistry JO - Curr Protoc Nucleic Acid Chem VL - 57 N2 - 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. SN - 1934-9289 UR - https://www.unboundmedicine.com/medline/citation/24961723/An_In_Vitro_Selection_Protocol_for_Threose_Nucleic_Acid__TNA__Using_DNA_Display_ L2 - https://dx.doi.org/10.1002/0471142700.nc0908s57 DB - PRIME DP - Unbound Medicine ER -