RNA self-splicing by engineered hairpin ribozyme variants

Small RNAs capable of self-cleavage and ligation might have been the precursors for the much more complex self-splicing group I and II introns in an early RNA world. Here, we demonstrate the activity of engineered hairpin ribozyme variants, which as self-splicing introns are removed from their parent RNA. In the process, two cleavage reactions are supported at the two intron-exon junctions, followed by ligation of the two generated exon fragments. As a result, the hairpin ribozyme, here acting as the self-splicing intron, is cut out. Two self-splicing hairpin ribozyme variants were investigated, one designed by hand, the other by a computer-aided approach. Both variants perform self-splicing, generating a cut-out intron and ligated exons.

Automated summary

This study demonstrates that small RNAs capable of self-cleavage and ligation may have been precursors for more complex self-splicing group I and II introns in an early RNA world. Engineered hairpin ribozyme variants, functioning as self-splicing introns, were shown to undergo two cleavage reactions at intron-exon junctions followed by ligation of exon fragments, leading to the removal of the ribozyme. Two self-splicing hairpin ribozyme variants, one designed manually and the other using a computer-aided approach, were found to efficiently perform self-splicing.