Bench-Top Fabrication of Single-Molecule Nanoarrays by DNA Origami Placement
Year: 2021
Type: article
Abstract: Large-scale nanoarrays of single biomolecules enable high-throughput assays while unmasking the underlying heterogeneity within ensemble populations. Until recently, creating such grids which combine the advantages of microarrays and single-molecule experiments (SMEs) has been particularly challenging due to the mismatch between the size of these molecules and the resolution of top-down fabrication techniques. DNA origami placement (DOP) combines two powerful techniques to address this issue: (i) DNA origami, which provides a ∼100 nm self-assembled template for single-molecule organization with 5 nm resolution and (ii) top-down lithography, which patterns these DNA nanostructures, transforming them into functional nanodevices via large-scale integration with arbitrary substrates. Presently, this technique relies on state-of-the-art infrastructure and highly trained personnel, making it prohibitively expensive for researchers. Here, we introduce a cleanroom-free, $1 benchtop technique to create meso-to-macro-scale DNA origami nanoarrays using self-assembled colloidal nanoparticles, thereby circumventing the need for top-down fabrication. We report a maximum yield of 74%, 2-fold higher than the statistical limit of 37% imposed on non-specific molecular loading alternatives. Furthermore, we provide a proof-of-principle for the ability of this nanoarray platform to transform traditionally low-throughput, stochastic, single-molecule assays into high-throughput, deterministic ones, without compromising data quality. Our approach has the potential to democratize single-molecule nanoarrays and demonstrates their utility as a tool for biophysical assays and diagnostics.
Source: ACS Nano
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $5,000
Funders: National Science Foundation, Arizona State University, Arizona Biomedical Research Commission, National Institutes of Health, Office of Naval Research, NIH Office of the Director, National Institute of Allergy and Infectious Diseases, Division of Computing and Communication Foundations, Division of Civil, Mechanical and Manufacturing Innovation, Office of Naval Research Global
Awards: G190486606, Collaborative Research: Molecular Programming Architectures, Abstractions, Algorithms, and Applications, G3720352588, G382725243, SemiSynBio-II: Programmable abiotic-biotic interface with planar DNA nanopore electrodes, G4504108201, G5271448993, G5829289835, Nanoscale reconstruction of mechanical systems involved in disease pathogenesis, G6101456529, G6142989802, Asymmetric DNA Shapes for Orienting Molecular Components within Hybrid Nanodevices, G779436476, G8376432556, G8876996369
OpenAlex ID: W3182061642