Journal
BIOCHEMISTRY
Volume 58, Issue 23, Pages 2675-2681Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.biochem.9b00063
Keywords
-
Categories
Funding
- European Research Council (ERC) under the European's Union Horizon 2020 program [770940]
- Deutsche Forschungsgemeinschaft [364775124]
- Ville de Paris Emergences program
- Marie Sklodowska-Curie fellowship from the European Union's Horizon 2020 program [795580]
- PRESTIGE grant from the European Union's Seventh Framework Programme [609102]
- Marie Curie Actions (MSCA) [795580] Funding Source: Marie Curie Actions (MSCA)
- European Research Council (ERC) [770940] Funding Source: European Research Council (ERC)
Ask authors/readers for more resources
In the absence of DNA, a solution containing the four deoxynucleotidetriphosphates (dNTPs), a DNA polymerase, and a nicking enzyme generates a self-replicating mixture of DNA species called parasite. Parasites are problematic in template-based isothermal amplification schemes such as EXPAR as well as in related molecular programming approaches, such as the PEN DNA toolbox. Here we show that using a nicking enzyme with only three letters (C, G, T) in the top strand of its recognition site, such as Nb.BssSI, allows us to change the sequence design of EXPAR templates in a way that prevents the formation of parasites when dATP is removed from the solution. This method allows us to make the EXPAR reaction robust to parasite contamination, a common feature in the laboratory, while keeping it compatible with PEN programs, which we demonstrate by engineering a parasite-proof bistable reaction network.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available