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Non-enzymatic primer extension with strand displacement

By Lijun Zhou, Chris Kim, Katherine H. Ho, Derek K. O’Flaherty, Constantin Giurgiu, Tom H Wright, Jack W Szostak

Posted 16 Sep 2019
bioRxiv DOI: 10.1101/770578 (published DOI: 10.7554/eLife.51888)

Non-enzymatic RNA self-replication is integral to the 'RNA World' hypothesis. Despite considerable progress in non-enzymatic template copying, true replication remains challenging due to the difficulty of separating the strands of the product duplex. Here, we report a prebiotically plausible solution to this problem in which short invader oligonucleotides unwind an RNA duplex through a toehold/branch migration mechanism, allowing non-enzymatic primer extension on a template that was previously occupied by its complementary strand. Kinetic studies of single-step reactions suggest that following invader binding, branch migration results in a 2:3 partition of the template between open and closed states. Finally, we demonstrate continued primer extension with strand displacement by employing activated 3′-aminonucleotides, a more reactive proxy for ribonucleotides. Our study suggests that complete cycles of non-enzymatic replication of the primordial genetic material may have been catalyzed by short RNA oligonucleotides.

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