The Synthesis of Ribozymes using TC-RNA Chemistry


Poster presented at IRT 2010 - XIX International Round Table on Nucleosides, Nucleotides and Nucleic Acids, Lyon, France.


Recently, 2’-O-thiomorpholine-4-carbothioate (TC) was introduced as an alternative protection chemistry to TBDMS in RNA synthesis. This utilises a one step simple deprotection method that removes both the 2’-TC and nucleobase protection at the same time. The 2’-TC protecting group has been designed such that it deprotects at a slower rate than the nucleobases, reducing the possibility of isomerisation during deprotection. Combined with the fact that the
coupling efficiency is comparable with DNA synthesis, there is the potential to synthesise good quality long RNA such as ribozymes. The synthesis of two ribozymes (55mer and 77mer) was carried out using TC-RNA phosphoramidite chemistry. Analysis of these was then carried out by MALDI MS, LCMS, and CGE. These were then compared to the same sequences synthesised via the more conventional TBDMS chemistry. The cleavage and ligation
activity of both sets of oligonucleotides were then evaluated.


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