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Issue 22, 2019
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Continuous nonenzymatic cross-replication of DNA strands with in situ activated DNA oligonucleotides

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Abstract

Continuous enzyme-free replication of oligonucleotides is central for open-ended evolution experiments that mimic the origin of life. Here, we studied a reaction system, whereby two 24mer DNA templates cross-catalyzed each other's synthesis from four 12mer DNA fragments, two of which were in situ activated with the condensing agent 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide (EDC). We circumvented the problem of product inhibition by melting the stable product duplexes for their reuse as templates in the following ligation step. The system reproduced itself through ligation/melting cycles and survived exponential dilution. We quantified EDC-induced side reactions in a detailed kinetic model. The model allowed us to analyze the effects of various reaction rates on the system's kinetics and confirmed maximal replication under the chosen conditions. The presented system enables us to study nonenzymatic open-ended evolution experiments starting from diverse sequence pools.

Graphical abstract: Continuous nonenzymatic cross-replication of DNA strands with in situ activated DNA oligonucleotides

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Supplementary files

Article information


Submitted
14 Feb 2019
Accepted
28 Apr 2019
First published
30 Apr 2019

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2019,10, 5807-5814
Article type
Edge Article

Continuous nonenzymatic cross-replication of DNA strands with in situ activated DNA oligonucleotides

E. Edeleva, A. Salditt, J. Stamp, P. Schwintek, J. Boekhoven and D. Braun, Chem. Sci., 2019, 10, 5807
DOI: 10.1039/C9SC00770A

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    [Original citation] - Published by The Royal Society of Chemistry.

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