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Enzymatic Synthesis of Nucleic Acids with Defined Regioisomeric 2'-5' Linkages.

ABSTRACT: Information-bearing nucleic acids display universal 3'-5' linkages, but regioisomeric 2'-5' linkages occur sporadically in non-enzymatic RNA synthesis and may have aided prebiotic RNA replication. Herein we report on the enzymatic synthesis of both DNA and RNA with site-specific 2'-5' linkages by an engineered polymerase using 3'-deoxy- or 3'-O-methyl-NTPs as substrates. We also report the reverse transcription of the resulting modified nucleic acids back to 3'-5' linked DNA with good fidelity. This enables a fast and simple method for "structural mutagenesis" by the position-selective incorporation of 2'-5' linkages, whereby nucleic acid structure and function may be probed through local distortion by regioisomeric linkages while maintaining the wild-type base sequence as we demonstrate for the 10-23?RNA endonuclease DNAzyme.

SUBMITTER: Cozens C

PROVIDER: S-EPMC4736440 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

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Enzymatic Synthesis of Nucleic Acids with Defined Regioisomeric 2'-5' Linkages.

Cozens Christopher C Mutschler Hannes H Nelson Geoffrey M GM Houlihan Gillian G Taylor Alexander I AI Holliger Philipp P

Angewandte Chemie (International ed. in English) 20151103 51

Information-bearing nucleic acids display universal 3'-5' linkages, but regioisomeric 2'-5' linkages occur sporadically in non-enzymatic RNA synthesis and may have aided prebiotic RNA replication. Herein we report on the enzymatic synthesis of both DNA and RNA with site-specific 2'-5' linkages by an engineered polymerase using 3'-deoxy- or 3'-O-methyl-NTPs as substrates. We also report the reverse transcription of the resulting modified nucleic acids back to 3'-5' linked DNA with good fidelity. ...[more]

PMID: 26527364

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