Efficient preparation of organic substrate-RNA conjugates via in vitro transcription.
A concise synthetic way has been developed for the preparation of guanosine monophosphate derivatives carrying a decaethylene glycol spacer at their 5'-oxygen to which are attached a range of organic substrates. The four different compounds, prepared via a convergent synthetic strategy, carry a tethered benzylallyl ether residue (1a), an anthracene (1b), a benzyl carbamate residue (1c), or a primary amino group (1d), respectively. All four compounds have been successfully incorporated at the 5'-end of a 25-mer long RNA transcript via T7 RNA polymerase, and no inhibition of chain elongation could be observed. Under proper conditions, 1a and 1b can be incorporated up to 90-95% and 1c up to 68%. The amino-terminated initiator 1d is incorporated less efficiently although still up to 49%. These results show that the more hydrophobic the guanosine monophosphate derivative is, the higher is its enzymatic incorporation.
Fiammengo
Roberto
R
Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany.
Musílek
Kamil
K
Jäschke
Andres
A
D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000378:metabolism; D006157:Guanosine Monophosphate / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D008968:Molecular Conformation; D012313:RNA / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism
D014764:Viral Proteins; D012313:RNA; D006157:Guanosine Monophosphate; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases