rEXPAR: An Isothermal Amplification Scheme That Is Robust to Autocatalytic Parasites - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Biochemistry Année : 2019

rEXPAR: An Isothermal Amplification Scheme That Is Robust to Autocatalytic Parasites

Résumé

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.
Fichier principal
Vignette du fichier
190417_no_parasite_biochemistry_accepted_version_for_arxiv.pdf (1.35 Mo) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)
Loading...

Dates et versions

hal-02180838 , version 1 (11-07-2019)

Identifiants

Citer

Georg Urtel, Marc van Der Hofstadt, Jean-Christophe Galas, André Estévez-Torres. rEXPAR: An Isothermal Amplification Scheme That Is Robust to Autocatalytic Parasites. Biochemistry, 2019, 58 (23), pp.2675-2681. ⟨10.1021/acs.biochem.9b00063⟩. ⟨hal-02180838⟩
73 Consultations
120 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More