Synthesis of modified nucleoside 3',5'-bisphosphates and their incorporation into oligoribonucleotides with T4 RNA ligase

Publisher Website
Google Scholar
Abstract
A simple procedure is described to prepare nucleoside 3''(2''),5''-bisphosphates from the corresponding nucleosides with the use of pyrophosphoryl chloride. This method is rapid, gives nearly quantitative yields and most importantly, can be used for a variety of nucleosides with base and sugar modifications. Since 3'',5''-bisphosphates are donors in the T4 RNA ligase reaction, a single residue can be enzymatically attached to the 3'' end of oligoribonucleotides. By these procedures, 5 different ring-modified nucleosides and 1 sugar-modified nucleoside were incorporated onto the 3'' end of (Ap)3C. In 2 cases, an additional step of synthesis with RNA ligase resulted in the modified nucleotide being located in an internal position in the oligonucleotide. Thus, a general method for the synthesis of oligoribonucleotides containing modified nucleosides is outlined. Since many of the modified nucleosides are fluorescent, oligomers containing them should be useful in a variety of physical and biochemical studies.