NanoBio&Med2018
november 20-22, 2018 - Barcelona (Spain) [5] Acuna, G.P. et al. Fluorescence enhancement at docking sites of DNA-directed self-assembled nanoantennas. Science 338, 506-510 (2012). [6] Nickels, P.C. et al. Molecular force spectroscopy with a DNA origami-based nanoscopic force clamp. Science 354, 305307 (2016)
DNA Origami Tools for NanoBioTec Philip Tinnefeld1, Viktorija Glembockyté, Lennart Grabenhorst, Ija Jusuk, Sarah Ochmann, Mario Raab, Tim Schröder, Florian Selbach, Florian Steiner, Johan Bohlen, Kristina Hübner, Jan Vogelsang, Andres Vera, Kateryna Trofymchuk1
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1Department
of Chemistry and Center for NanoScience, Ludwig-Maximilians-Universität München, 80539 München, Germany Philip.tinnefeld@lmu.de
Abstract In recent years, DNA nanotechnology has matured to enable robust production of complex nanostructures and hybrid materials. We have combined DNA nanotechnology with sensitive optical detection to create functional single-molecule devices that enable new applications in singlemolecule biophysics. Starting with superresolution nanorulers [1], a single-molecule mirage [2] and energy transfer switches [3] we developed DNA origami nano-adapters for targeted placement of single molecules in zeromode waveguides used for DNA sequencing [4]. Furthermore, a plasmonic fluorescence amplifier [5] is used for sensitive biosensing and single-molecule detection on low-tec detection devices such as a smartphone. The optical antennas assembled by DNA origami are further developed to improve fluorescence detection in molecular diagnostics. Finally, we present a molecular force spectroscopy employing DNA origami force clamps that work autonomously without any physical connection to the macroscopic world [6]. We used the conformer switching of a Holliday junction as a benchmark and studied the interaction of DNA binding proteins with DNA when the DNA is under 0-15 pN tension.
Figure 1. Sketch of a biomolecular sandwich assay of pathogenic DNA using signal amplification by a selfassembled DNA origami optical antenna.
References [1] Schmied, J.J. et al. DNA origami-based standards for quantitative fluorescence microscopy. Nature protocols 9, 1367-1391 (2014). [2] Raab, M., Vietz, C., Stefani, F.D., Acuna, G.P. & Tinnefeld, P. Shifting molecular localization by plasmonic coupling in a singlemolecule mirage. Nature communications 8, 13966 (2017). [3] Stein, I.H., Steinhauer, C. & Tinnefeld, P. SingleMolecule Four-Color FRET Visualizes EnergyTransfer Paths on DNA Origami. J Am Chem Soc 133, 4193-4195 (2011). [4] Pibiri, E., Holzmeister, P., Lalkens, B., Acuna, G.P. & Tinnefeld, P. Single-molecule positioning in zeromode waveguides by DNA origami nanoadapters. Nano letters 14, 3499-3503 (2014).
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