Synthesis of 5'-thiamine-capped RNA

• Verfasst von: Möhler, Marvin [VerfasserIn]
• Verfasst von: Höfer, Katharina [VerfasserIn]
• Verfasst von: Jäschke, Andres [VerfasserIn]
• Titel: Synthesis of 5'-thiamine-capped RNA
• Verf.angabe: Marvin Möhler, Katharina Höfer and Andres Jäschke
• E-Jahr: 2020
• Jahr: 24 November 2020
• Umfang: 16 S.
• Fussnoten: Gesehen am 25.01.2021
• Titel Quelle: Enthalten in: Molecules
• Ort Quelle: Basel : MDPI, 1996
• Jahr Quelle: 2020
• Band/Heft Quelle: 25(2020,23) Artikel-Nummer 5492, 16 Seiten
• ISSN Quelle: 1420-3049
• DOI: doi:10.3390/molecules25235492
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: chemical capture of thiamine-capped RNA
• Sach-SW: click chemistry
• Sach-SW: in vitro transcription
• Sach-SW: non-canonical initiating nucleotide (NCIN)
• Sach-SW: RNA modification
• Sach-SW: thiamine (vitamin B1)
• Sach-SW: thiamine adenosine triphosphate
• Sach-SW: thiamine-adenosine diphosphate
• Sach-SW: thiamine-capped RNA
• K10plus-PPN: 1745539964

Abstract

RNA 5'-modifications are known to extend the functional spectrum of ribonucleotides. In recent years, numerous non-canonical 5'-modifications, including adenosine-containing cofactors from the group of B vitamins, have been confirmed in all kingdoms of life. The structural component of thiamine adenosine triphosphate (thiamine-ATP), a vitamin B1 derivative found to accumulate in Escherichia coli and other organisms in response to metabolic stress conditions, suggests an analogous function as a 5'-modification of RNA. Here, we report the synthesis of thiamine adenosine dinucleotides and the preparation of pure 5'-thiamine-capped RNAs based on phosphorimidazolide chemistry. Furthermore, we present the incorporation of thiamine-ATP and thiamine adenosine diphosphate (thiamine-ADP) as 5'-caps of RNA by T7 RNA polymerase. Transcripts containing the thiamine modification were modified specifically with biotin via a combination of thiazole ring opening, nucleophilic substitution and copper-catalyzed azide-alkyne cycloaddition. The highlighted methods provide easy access to 5'-thiamine RNA, which may be applied in the development of thiamine-specific RNA capture protocols as well as the discovery and confirmation of 5'-thiamine-capped RNAs in various organisms.