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Nonenzymatic loop-closing ligation generates RNA hairpins and is a template-free way to assemble functional RNAs

Wu, L.-F.; Liu, Z.; Roberts, S. J.; Su, M.; Szostak, J. W.; Sutherland, J. D.

2021-11-19 synthetic biology
10.1101/2021.11.19.469337 bioRxiv
Show abstract

RNA hairpin loops are the predominant element of secondary structure in functional RNAs. The emergence of primordial functional RNAs, such as ribozymes that fold into complex structures that contain multiple hairpin loops, is generally thought to have been supported by template-directed ligation. However, template inhibition and RNA misfolding problems impede the emergence of function. Here we demonstrate that RNA hairpin loops can be synthesized directly from short RNA duplexes with single-stranded overhangs by nonenzymatic loop-closing ligation chemistry. We show that loop-closing ligation allows full-length functional ribozymes containing a hairpin loop to be assembled free of inhibitory template strands. This approach to the assembly of structurally complex RNAs suggests a plausible pathway for the emergence of functional RNAs before a full-length RNA copying process became available.

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