Spontaneous Covalent Self-Assembly of the Azoarcus Ribozyme from Five Fragments

Spontaneous covalent assembly of short RNA fragments has been proposed as a plausible prebiotically relevant pathway to a self-reproducing system. We previously showed that the Azoarcus group I intron could self-assemble from four RNA fragments. Here, we extended this fragmentation to five RNAs that averaged <40 nucleotides in length. We optimized this reaction and showed that a dehydration-rehydration sequence was the most effective means to date to shift the self-assembly equilibrium from reactants to products.