Enzymatic synthesis of base-modified RNA by T7 RNA polymerase. A systematic study and comparison of 5-substituted pyrimidine and 7-substituted 7-deazapurine nucleoside triphosphates as substrates
Jazyk angličtina Země Anglie, Velká Británie Médium print
Typ dokumentu srovnávací studie, časopisecké články, práce podpořená grantem
PubMed
30063056
DOI
10.1039/c8ob01498a
Knihovny.cz E-zdroje
- MeSH
- adenosintrifosfát analogy a deriváty metabolismus MeSH
- bakteriofág T7 enzymologie MeSH
- cytidintrifosfát analogy a deriváty metabolismus MeSH
- DNA řízené RNA-polymerasy metabolismus MeSH
- purinové nukleosidy chemie metabolismus MeSH
- puriny chemie metabolismus MeSH
- pyrimidinové nukleosidy chemie metabolismus MeSH
- RNA chemie metabolismus MeSH
- substrátová specifita MeSH
- uridintrifosfát analogy a deriváty metabolismus MeSH
- virové proteiny metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- 7-deazapurine MeSH Prohlížeč
- adenosintrifosfát MeSH
- bacteriophage T7 RNA polymerase MeSH Prohlížeč
- cytidintrifosfát MeSH
- DNA řízené RNA-polymerasy MeSH
- purinové nukleosidy MeSH
- puriny MeSH
- pyrimidinové nukleosidy MeSH
- RNA MeSH
- uridintrifosfát MeSH
- virové proteiny MeSH
We synthesized a small library of eighteen 5-substituted pyrimidine or 7-substituted 7-deazapurine nucleoside triphosphates bearing methyl, ethynyl, phenyl, benzofuryl or dibenzofuryl groups through cross-coupling reactions of nucleosides followed by triphosphorylation or through direct cross-coupling reactions of halogenated nucleoside triphosphates. We systematically studied the influence of the modification on the efficiency of T7 RNA polymerase catalyzed synthesis of modified RNA and found that modified ATP, UTP and CTP analogues bearing smaller modifications were good substrates and building blocks for the RNA synthesis even in difficult sequences incorporating multiple modified nucleotides. Bulky dibenzofuryl derivatives of ATP and GTP were not substrates for the RNA polymerase. In the case of modified GTP analogues, a modified procedure using a special promoter and GMP as initiator needed to be used to obtain efficient RNA synthesis. The T7 RNA polymerase synthesis of modified RNA can be very efficiently used for synthesis of modified RNA but the method has constraints in the sequence of the first three nucleotides of the transcript, which must contain a non-modified G in the +1 position.
Citace poskytuje Crossref.org
Synthesis and Biological Profiling of Quinolino-Fused 7-Deazapurine Nucleosides