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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2019
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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. |
format | Online Article Text |
id | pubmed-6872209 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
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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