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Non-enzymatic primer extension with strand displacement

Non-enzymatic RNA self-replication is integral to the emergence of the ‘RNA World’. Despite considerable progress in non-enzymatic template copying, demonstrating a full replication cycle remains challenging due to the difficulty of separating the strands of the product duplex. Here, we report a pre...

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Detalles Bibliográficos
Autores principales: Zhou, Lijun, Kim, Seohyun Chris, Ho, Katherine H, O'Flaherty, Derek K, Giurgiu, Constantin, Wright, Tom H, Szostak, Jack W
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6872209/
https://www.ncbi.nlm.nih.gov/pubmed/31702557
http://dx.doi.org/10.7554/eLife.51888
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author Zhou, Lijun
Kim, Seohyun Chris
Ho, Katherine H
O'Flaherty, Derek K
Giurgiu, Constantin
Wright, Tom H
Szostak, Jack W
author_facet Zhou, Lijun
Kim, Seohyun Chris
Ho, Katherine H
O'Flaherty, Derek K
Giurgiu, Constantin
Wright, Tom H
Szostak, Jack W
author_sort Zhou, Lijun
collection PubMed
description Non-enzymatic RNA self-replication is integral to the emergence of the ‘RNA World’. Despite considerable progress in non-enzymatic template copying, demonstrating a full replication cycle remains challenging due to the difficulty of separating the strands of the product duplex. Here, we report a prebiotically plausible approach to strand displacement synthesis 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 facilitated by short RNA oligonucleotides.
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spelling pubmed-68722092019-11-23 Non-enzymatic primer extension with strand displacement Zhou, Lijun Kim, Seohyun Chris Ho, Katherine H O'Flaherty, Derek K Giurgiu, Constantin Wright, Tom H Szostak, Jack W eLife Biochemistry and Chemical Biology Non-enzymatic RNA self-replication is integral to the emergence of the ‘RNA World’. Despite considerable progress in non-enzymatic template copying, demonstrating a full replication cycle remains challenging due to the difficulty of separating the strands of the product duplex. Here, we report a prebiotically plausible approach to strand displacement synthesis 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 facilitated by short RNA oligonucleotides. eLife Sciences Publications, Ltd 2019-11-08 /pmc/articles/PMC6872209/ /pubmed/31702557 http://dx.doi.org/10.7554/eLife.51888 Text en © 2019, Zhou et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biochemistry and Chemical Biology
Zhou, Lijun
Kim, Seohyun Chris
Ho, Katherine H
O'Flaherty, Derek K
Giurgiu, Constantin
Wright, Tom H
Szostak, Jack W
Non-enzymatic primer extension with strand displacement
title Non-enzymatic primer extension with strand displacement
title_full Non-enzymatic primer extension with strand displacement
title_fullStr Non-enzymatic primer extension with strand displacement
title_full_unstemmed Non-enzymatic primer extension with strand displacement
title_short Non-enzymatic primer extension with strand displacement
title_sort non-enzymatic primer extension with strand displacement
topic Biochemistry and Chemical Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6872209/
https://www.ncbi.nlm.nih.gov/pubmed/31702557
http://dx.doi.org/10.7554/eLife.51888
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