A multifunctional DNA nanostructure based on multicolor FRET for nuclease activity assay. Issue 18 (11th August 2020)

• Record Type: Journal Article
• Authors: Hu, Juan; Li, Wen-can; Qiu, Jian-Ge; Jiang, BingHua; Zhang, Chun-yang
• Is Part Of: Analyst. Volume 145:Issue 18(2020)
• Page Start: 6054
• Page End: 6060
• Publication Date: 2020-08-11
• Subjects: Chemistry, Analytic -- Periodicals; 543
• DOI: 10.1039/d0an01212b
• Languages: English
• ISSNs: 0003-2654

Abstract

Abstract : We develop a four-color fluorescent probe for ratiometric detection of multiple nucleases based on multistep fluorescence resonance energy transfer. Abstract : Nucleases play a crucial role in DNA replication, recombination and repair which are associated with cancers. Herein, we develop a four-color fluorescent probe for ratiometric detection of multiple nucleases. This four-color fluorescent probe consists of four fluorescent dyes connected by a DNA tetrahedral nanostructure with the involvement of multistep fluorescence resonance energy transfer (FRET). Based on the principle of self-assembly, the four-color fluorescent probe is constructed by integrating one acceptor with three spatially and spectrally distinct acceptors. A DNA tetrahedral nanostructure functions as a scaffold to link the acceptor dyes ( i.e., diethylaminocoumarin (DEA), carboxyfluorescein (FAM), Texas Red, and Cy5). The fluorescence emissions of DEA, FAM, Texas Red and Cy5 can be observed through efficient multi-step energy transfer. This four-color fluorescent probe enables single excitation/four emissions, and it can be used for ratiometric detection of nucleases ( i.e., XhoI, HindIII and KpnI) and the screening of nuclease inhibitors. Importantly, this four-color fluorescent probe can be further applied to discriminate multiple biomolecule targets by simply integrating the recognition sites of various biomolecules into the DNA tetrahedral nanostructure.