Tuning DNA–nanoparticle conjugate properties allows modulation of nuclease activity. Issue 9 (25th February 2021)

• Record Type: Journal Article
• Authors: Hsiao, Jeff C.; Buryska, Tomas; Kim, Eunjung; Howes, Philip D.; deMello, Andrew J.
• Is Part Of: Nanoscale. Volume 13:Issue 9(2021)
• Page Start: 4956
• Page End: 4970
• Publication Date: 2021-02-25
• Subjects: Nanoscience -- Periodicals; Nanotechnology -- Periodicals; 620.505
• DOI: 10.1039/d0nr08668a
• Languages: English
• ISSNs: 2040-3364

Abstract

Abstract : A systematic study of the interactions between nucleases and oligonucleotide-coated gold nanoparticles (spherical nucleic acids, SNAs) demonstrates that the modular units of SNAs can be leveraged to either accelerate or suppress nuclease kinetics. Abstract : Enzyme–nanoparticle interactions can give rise to a range of new phenomena, most notably significant enzymatic rate enhancement. Accordingly, the careful study and optimization of such systems is likely to give rise to advanced biosensing applications. Herein, we report a systematic study of the interactions between nuclease enzymes and oligonucleotide-coated gold nanoparticles (spherical nucleic acids, SNAs), with the aim of revealing phenomena worthy of evolution into functional nanosystems. Specifically, we study two nucleases, an exonuclease (ExoIII) and an endonuclease (Nt.BspQI), via fluorescence-based kinetic experiments, varying parameters including enzyme and substrate concentrations, and nanoparticle size and surface coverage in non-recycling and a recycling formats. We demonstrate the tuning of nuclease activity by SNA characteristics and show that the modular units of SNAs can be leveraged to either accelerate or suppress nuclease kinetics. Additionally, we observe that the enzymes are capable of cleaving restriction sites buried deep in the oligonucleotide surface layer and that enzymatic rate enhancement occurs in the target recycling format but not in the non-recycling format. Furthermore, we demonstrate a new SNA phenomenon, we term 'target stacking', whereby nucleic acid hybridization efficiency increases as enzyme cleavage proceeds during the beginning of a reaction. This investigation provides important data to guide the design of novel SNAs in biosensing and in vitro diagnostic applications.