Synthesis, Dynamic Combinatorial Chemistry, and PCR Amplification of 3′-5′ and 3′-6′ Disulfide-linked Oligonucleotides

• Published in: Angewandte Chemie Vol. 126; no. 52; pp. 14643 - 14646
• Main Authors: Hansen, Dennis Jul, Manuguerra, Ilenia, Kjelstrup, Michael Brøndum, Gothelf, Kurt Vesterager
• Format: Journal Article
• Language: English; German
• Published: Weinheim WILEY-VCH Verlag 22.12.2014; WILEY‐VCH Verlag; Wiley Subscription Services, Inc
• Subjects: Amplification; Chemistry; Combinatorial chemistry; Disulfide; Disulfides; Dynamics; Dynamische kombinatorische Chemie; Festphasensynthese; Links; Mutations; Nucleinsäuren; Oligonucleotides; Primer-Extension; Sequencing; Synthesis
• ISSN: 0044-8249; 1521-3757
• DOI: 10.1002/ange.201405761

Disulfide dithymidines linked 3′–5′ or 3′–6′ were synthesized and incorporated into oligonucleotides through a combined phosphotriester and phosphoramidite solid‐phase oligonucleotide synthesis approach. The disulfide links are cleaved and formed reversibly in the presence of thiols and oligonucleotides. This link was shown to be sequence‐adaptive in response to given templates in the presence of mercaptoethanol. The artificial 3′–5′ and 3′–6′ disulfide link was tolerated by polymerases in the polymerase chain reaction (PCR). By using sequencing analysis, we show that single mutations frequently occurred randomly in the amplification products of the PCR. 3′–5′‐ und 3′–6′‐Internucleosid‐Disulfidbindungen ermöglichen die Spaltung und templatgesteuerte Bildung gewünschter Disulfide in Gegenwart von Mercaptoethanol. Das künstliche Disulfidrückgrat wird von Polymerasen toleriert, und die Sequenzen können mithilfe der Polymerase‐Kettenreaktion amplifiziert werden.