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
Language English Country England, Great Britain Media print
Document type Comparative Study, Journal Article, Research Support, Non-U.S. Gov't
PubMed
30063056
DOI
10.1039/c8ob01498a
Knihovny.cz E-resources
- MeSH
- Adenosine Triphosphate analogs & derivatives metabolism MeSH
- Bacteriophage T7 enzymology MeSH
- Cytidine Triphosphate analogs & derivatives metabolism MeSH
- DNA-Directed RNA Polymerases metabolism MeSH
- Purine Nucleosides chemistry metabolism MeSH
- Purines chemistry metabolism MeSH
- Pyrimidine Nucleosides chemistry metabolism MeSH
- RNA chemistry metabolism MeSH
- Substrate Specificity MeSH
- Uridine Triphosphate analogs & derivatives metabolism MeSH
- Viral Proteins metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
- Names of Substances
- 7-deazapurine MeSH Browser
- Adenosine Triphosphate MeSH
- bacteriophage T7 RNA polymerase MeSH Browser
- Cytidine Triphosphate MeSH
- DNA-Directed RNA Polymerases MeSH
- Purine Nucleosides MeSH
- Purines MeSH
- Pyrimidine Nucleosides MeSH
- RNA MeSH
- Uridine Triphosphate MeSH
- Viral Proteins 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.
References provided by Crossref.org
Synthesis and Biological Profiling of Quinolino-Fused 7-Deazapurine Nucleosides