Enzymatic Synthesis of Nucleic Acids with Defined Regioisomeric 2′‐5′ Linkages. Issue 51 (3rd November 2015)

• Record Type: Journal Article
• Authors: Cozens, Christopher; Mutschler, Hannes; Nelson, Geoffrey M.; Houlihan, Gillian; Taylor, Alexander I.; Holliger, Philipp
• Is Part Of: Angewandte Chemie international edition. Volume 54:Issue 51(2015)
• Page Start: 15570
• Page End: 15573
• Publication Date: 2015-11-03
• Subjects: DNAzyme -- nucleic acids -- nucleotides -- polymerase -- regioselectivity; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201508678
• Languages: English
• ISSNs: 1433-7851

Abstract

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. Abstract : Specific distortion : A novel engineered polymerase can synthesize both DNA and RNA with regioisomeric 2′‐5′ backbone linkages. It forms the basis for position‐selective incorporation of 2′‐5′ linkages enabling "structural mutagenesis". As a result structure, conformation, duplex stability, and activity of nucleic acids may be probed through the site‐specific insertion of regioisomeric backbone distortions.