title: Modular Nuclease-Responsive DNA Three-Way Junction-Based Dynamic Assembly of a DNA Device and Its Sensing Application.
abstract: Here, we explored a modular strategy for rational design of nuclease-responsive three-way junctions (TWJs) and fabricated a dynamic DNA device in a "plug-and-play" fashion. First, inactivated TWJs were designed, which contained three functional domains: the inaccessible toehold and branch migration domains, the specific sites of nucleases, and the auxiliary complementary sequence. The actions of different nucleases on their specific sites in TWJs caused the close proximity of the same toehold and branch migration domains, resulting in the activation of the TWJs and the formation of a universal trigger for the subsequent dynamic assembly. Second, two hairpins (H1 and H2) were introduced, which could coexist in a metastable state, initially to act as the components for the dynamic assembly. Once the trigger initiated the opening of H1 via TWJs-driven strand displacement, the cascade hybridization of hairpins immediately switched on, resulting in the formation of the concatemers of H1/H2 complex appending numerous integrated G-quadruplexes, which were used to obtain label-free signal readout. The inherent modularity of this design allowed us to fabricate a flexible DNA dynamic device and detect multiple nucleases through altering the recognition pattern slightly. Taking uracil-DNA glycosylase and CpG methyltransferase M.SssI as models, we successfully realized the butt joint between the uracil-DNA glycosylase and M.SssI recognition events and the dynamic assembly process. Furthermore, we achieved ultrasensitive assay of nuclease activity and the inhibitor screening. The DNA device proposed here will offer an adaptive and flexible tool for clinical diagnosis and anticancer drug discovery.
authors.affiliation: Key Laboratory for Colloid and Interface Chemistry of Education Ministry, School of Chemistry and Chemical Engineering, Shandong University , 250100 Jinan, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.affiliation: School of Pharmaceutical Sciences, Shandong University , 250012 Jinan, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Key Laboratory for Colloid and Interface Chemistry of Education Ministry, School of Chemistry and Chemical Engineering, Shandong University , 250100 Jinan, People's Republic of China.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Xu
authors.affiliation: Key Laboratory for Colloid and Interface Chemistry of Education Ministry, School of Chemistry and Chemical Engineering, Shandong University , 250100 Jinan, People's Republic of China.
authors.forename: Haiping
authors.initials: H
authors.lastname: Wei
authors.affiliation: Key Laboratory for Colloid and Interface Chemistry of Education Ministry, School of Chemistry and Chemical Engineering, Shandong University , 250100 Jinan, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Jiang
mesh_terms: D000970:Antineoplastic Agents / Q000032:analysis / Q000494:pharmacology; D015374:Biosensing Techniques; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D054856:G-Quadruplexes; D006801:Humans; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D000970:Antineoplastic Agents; D004247:DNA; D003851:Deoxyribonucleases