2-Selenouridine triphosphate synthesis and Se-RNA transcription

2-Selenouridine (^SeU) is one of the naturally occurring modifications of Se-tRNAs (^SeU-RNA) at the wobble position of the anticodon loop. Its role in the RNA-RNA interaction, especially during the mRNA decoding, is elusive. To assist the research exploration, herein we report the enzymatic synthesis of the ^SeU-RNA via 2-selenouridine triphosphate (^SeUTP) synthesis and RNA transcription. Moreover, we have demonstrated that the synthesized ^SeUTP is stable and recognizable by T7 RNA polymerase. Under the optimized conditions, the transcription yield of ^SeU-RNA can reach up to 85% of the corresponding native RNA. Furthermore, the transcribed ^SeU-hammerhead ribozyme has the similar activity as the corresponding native, which suggests usefulness of ^SeU-RNAs in function and structure studies of noncoding RNAs, including the Se-tRNAs.