Regensburg 2019 – wissenschaftliches Programm
SYDN 1.4: Hauptvortrag
Donnerstag, 4. April 2019, 11:15–11:45, H1
DNA origami nanostructures aid the super-resolution microscopy interrogation of proteins and allow single-molecule force measurements on biological systems — Leonhard Jakob1, Kevin Kramm1, Julia Molle2, Tim Schröder2, Philip Nickels3, Alessandro Vannini4, Tim Liedl3, Philip Tinnefeld2, and •Dina Grohmann1 — 1Department of Biochemistry, Genetics and Microbiology, Institute of Microbiology, Single-Molecule Biochemistry Lab, University of Regensburg, Universitätsstraße 31, 93053 Regensburg, Germany — 2Department for Chemistry and Center for Nanoscience, Ludwig-Maximilians-Universität München, Butenandtstr. 11, 81377 München, Germany — 3Faculty of Physics & Center for Nanoscience (CeNS), Ludwig-Maximilians-Universität (LMU), Geschwister-Scholl-Platz 1, 80539 München, Germany — 4The Institute of Cancer Research, London SW7 3RP, UK
DNA nanotechnology is aiming to create complex functional structures on the nanometre scale with the aim, among others, to exploit these structures in biological research. While DNA-scaffolded nanostructures have been successfully developed over the last decade, the integration of functional biological molecules into DNA origami structures remains challenging. Here, we report on two projects that combine the DNA origami technique with single-molecule microscopy/spectroscopy to gain insights into the structure and function of protein assemblies.
(1) A long-standing question has been whether super-resolution (SR) microscopy can be employed for the structural interrogation of proteins in the sub-20 nm range. Here, we show that the marriage of DNA nanotechnology and single-molecule biochemistry allows the first steps towards the investigation of the structural organization of a protein using DNA PAINT SR microscopy.
(2) We exploit a self-assembled molecular force clamp built from DNA to perform force spectroscopy measurements on biological samples. Among others, we probed the force sensitivity of the DNA scissor Cas9 and quantified the influence of DNA strain on the assembly of the transcription initiation machinery.