Cardiff University | Prifysgol Caerdydd ORCA
Online Research @ Cardiff 
WelshClear Cookie - decide language by browser settings

Single molecule DNA origami nanoarrays with controlled protein orientation

Cervantes-Salguero, K., Freeley, M., Gwyther, R. E. A., Jones, D. D. ORCID:, Chavez, J. L. and Palma, M. 2022. Single molecule DNA origami nanoarrays with controlled protein orientation. Biophysics Reviews 3 (3) 10.1063/5.0099294

[thumbnail of Revised-SuppMat_single protein nanoarrays.pdf] PDF - Supplemental Material
Available under License Creative Commons Attribution.

Download (562kB)
[thumbnail of Revised-Manuscript_single protein nanoarrays_.pdf] PDF - Accepted Post-Print Version
Available under License Creative Commons Attribution.

Download (772kB)


The nanoscale organization of functional (bio)molecules on solid substrates with nanoscale spatial resolution and single-molecule control—in both position and orientation—is of great interest for the development of next-generation (bio)molecular devices and assays. Herein, we report the fabrication of nanoarrays of individual proteins (and dyes) via the selective organization of DNA origami on nanopatterned surfaces and with controlled protein orientation. Nanoapertures in metal-coated glass substrates were patterned using focused ion beam lithography; 88% of the nanoapertures allowed immobilization of functionalized DNA origami structures. Photobleaching experiments of dye-functionalized DNA nanostructures indicated that 85% of the nanoapertures contain a single origami unit, with only 3% exhibiting double occupancy. Using a reprogrammed genetic code to engineer into a protein new chemistry to allow residue-specific linkage to an addressable ssDNA unit, we assembled orientation-controlled proteins functionalized to DNA origami structures; these were then organized in the arrays and exhibited single molecule traces. This strategy is of general applicability for the investigation of biomolecular events with single-molecule resolution in defined nanoarrays configurations and with orientational control of the (bio)molecule of interest.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Publisher: AIP
ISSN: 2688-4089
Date of First Compliant Deposit: 21 July 2022
Date of Acceptance: 20 July 2022
Last Modified: 10 Oct 2023 19:06

Actions (repository staff only)

Edit Item Edit Item


Downloads per month over past year

View more statistics