Advanced Single Molecule Fluorescence Spectroscopy to analyze RNA and protein dynamics (P16)

• Subject Area: Biophysics
• Term: since 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 469281184

The project has two core methodological goals. Firstly, the development of an active single-molecule tracking/trapping method that will enable to perform fluorescence spectroscopy (Förster resonance energy transfer, i.e., FRET) on freely diffusing single molecules for a much longer time than currently possible. This shall be done by combining a so-called Anti-Brownian Electrophoretic (ABEL) trap with nanofluidics. Secondly, the development of a regenerating fluorescence labeling technique that circumvents the current limitations of single-molecule fluorescence spectroscopy due to photobleaching. This shall be achieved by adopting the method of label exchange as used in the super-resolution microscopy method of Points Accumulation for Imaging in Nanoscale Topography (PAINT). Both these advances, ABEL trap and label exchange, will open the door for single-molecule spectroscopy of biomolecules without time limitation and thus will enable us to observe and study rare conformational states, rare interaction events, and slow temporal biomolecular dynamics.

• DFG Programme: Collaborative Research Centres
• Subproject of: SFB 1565: Molecular mechanisms and interplay of gene expression processes
• Applicant Institution: Georg-August-Universität Göttingen
• Project Head: Professor Dr. Jörg Enderlein