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

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 activ...

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Published inChemical science (Cambridge) Vol. 10; no. 22; pp. 5807 - 5814
Main Authors Edeleva, Evgeniia, Salditt, Annalena, Stamp, Julian, Schwintek, Philipp, Boekhoven, Job, Braun, Dieter
Format Journal Article
LanguageEnglish
Published England Royal Society of Chemistry 14.06.2019
Subjects
Chemical synthesis
Computer simulation
Deoxyribonucleic acid
Dilution
DNA
Evolution
Oligonucleotides
Product inhibition
Reaction kinetics
Replication
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Summary: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 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.
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ISSN:2041-6520
2041-6539
DOI:10.1039/c9sc00770a