@article {38276,
	title = {Engineering DNA Nanocube SAM Scaffolds for FRET-Based Biosensing: Interfacial Characterization and Sensor Demonstration},
	journal = {Journal of the American Chemical Society},
	volume = {146},
	year = {2024},
	pages = {31560{\textendash}31573},
	abstract = {<p>Decorating a gold surface with molecular-level control over the positioning of DNA probes was demonstrated using a self-assembled monolayer (SAM) of wireframe DNA nanocube structures. The DNA nanocubes were specifically adsorbed and oriented using thiol-modified DNA on one face of the cube. The DNA nanocube SAM had a uniform coverage over the gold single crystal bead electrode with a separation of 20\&ndash;30 nm measured by AFM. The face of the nanocube furthest from the gold surface was designed to hybridize with two different sequences of a 50 base single-stranded DNA probe that was modified with a fluorophore. The first 20 bases were hybridized with the DNA nanocube. One of a pair of FRET fluorophores was used for each probe strand. The dimensions of the nanocube controlled the relative spacing between these fluorophores. When the DNA probes were single-stranded, a FRET signal was observed. FRET decreased to background levels when a complementary DNA target was hybridized to either probe, resulting in a turn-off sensor with little cross-talk between the individual hybridization events. Hybridization isotherms for one target gave K A = 170 pM and a detection limit \&lt;50 pM. In addition, the DNA nanocube SAM was configured to be used as a turn-on NeutrAvidin sensor using biotinylated DNA targets hybridized to each probe resulting in an increase in FRET. We show that the wireframe DNA nanocube can be an effective scaffold for preparing biosensors with controlled separation between surface-bound probes facilitating precise sensor surface design and enabling a wide range of sensing modalities with more than one signal available for correlative confirmation of the target binding.</p>},
	issn = {0002-7863},
	doi = {10.1021/jacs.4c09240},
	author = {Grzedowski, Adrian Jan and Jun, Daniel and Mahey, Amita and Zhou, Geyang Chris and Fernandez, Rachel and Bizzotto, Dan}
}
