Abstract

The topic of my presentation will be how a combination of protein engineering and synthetic chemistry can be exploited to generate fluorescent and bioluminescent probes for live-cell imaging. In the first part of my presentation I will introduce a highly permeable and biocompatible near-infrared fluorophore that can be specifically coupled to intracellular proteins in live cells and tissues using different labeling techniques. The fluorogenic character of the probe and its high brightness permit live-cell imaging experiments without washing steps. The fluorophore is ideally suited for live-cell superresolution microscopy approaches using either stimulated emission depletion (STED) or stochastic single-molecule localization nanoscopy. The excellent spectroscopic properties of the probe combined with its ease of use in live-cell applications make it a powerful new tool for bioimaging. In the second part of my talk, I will talk about our attempts to introduce a new class of fluorescent sensor proteins that permit to visualize drug and metabolite concentrations in living cells with high spatial and temporal resolution. I will also discuss how these sensor proteins can be utilized for point-of-care therapeutic drug monitoring.

Relevant papers:

1) “Fluorogenic probes for live-cell imaging of the cytoskeleton” Lukinavicius, G.; Reymond, L.; D’Este, E.; Masharina, A.; Gottfert, F.; Ta, H.; Guther, A.; Fournier, M.; Rizzo, S.; Waldmann, H.; Blaukopf, C.; Sommer, C.; Gerlich, D. W.; Arndt, H. D.; Hell, S. W.; Johnsson, K. Nature Methods 2014, 11, 731 2) “Bioluminescent sensor proteins for point-of-care therapeutic drug monitoring” Griss, R.; Schena, A.; Reymond, L.; Patiny, L.; Werner, D.; Tinberg, C. E.; Baker, D.; Johnsson, K. Nature Chem Biol 2014, 10, 598 3) “A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins” Lukinavicius, G.; Umezawa, K.; Olivier, N.; Honigmann, A.; Yang, G.; Plass, T.; Mueller, V.; Reymond, L.; Correa, I. R., Jr.; Luo, Z. G.; Schultz, C.; Lemke, E. A.; Heppenstall, P.; Eggeling, C.; Manley, S.; Johnsson, K. Nature Chemistry 2013, 5, 132