Bench-Top Fabrication of Single-Molecule Nanoarrays by DNA Origami Placement
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acsnano.1c01150.pdf
Description
Published version
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9.37 MB
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Unknown
Checksum (MD5)
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Author(s)
Shetty, Rishabh M.
Brady, Sarah R.
Rothemund, Paul W. K.
Hariadi, Rizal F.
Gopinath, Ashwin
Date Issued
July 2021
Journal
ACS Nano
Publisher
American Chemical Society (ACS)
Citation
Shetty, Rishabh M, Brady, Sarah R, Rothemund, Paul WK, Hariadi, Rizal F and Gopinath, Ashwin. 2021. "Bench-Top Fabrication of Single-Molecule Nanoarrays by DNA Origami Placement." ACS Nano, 15 (7).
Version
Final published version
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.
MIT Department
Massachusetts Institute of Technology. Department of Mechanical Engineering
Terms of Use
Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.1021/acsnano.1c01150
