Virtual International Mammalian Synthetic Biology Workshop (mSBW) 2020

Rapid Directed Molecular Evolution of Fluorescent Proteins in Mammalian Cells

Authors

Erica Jung, University of Illinois at Chicago
Kazuhiko Namikawa, Technische Universität Braunschweig
Oksana Subach, National Research Center “Kurchatov Institute”
Dmitry Korzhenevskiy, National Research Center “Kurchatov Institute”
Tatiana Rakitina, National Research Center “Kurchatov Institute”
Xian Xiao, Westlake University
Danielle Cosio, Media Lab, Massachusetts Institute of Technology
Jing Shi, Westlake Laboratory of Life Sciences and Biomedicine, Westlake University
Shaofeng Wu, Westlake Laboratory of Life Sciences and Biomedicine, Westlake University
Demian Park, Media Lab, Massachusetts Institute of Technology
Lea Eisenhard, Division of Cellular and Molecular Neurobiology, Zoological Institute, Technische Universität Braunschweig
Hongyun Tang, Westlake Laboratory of Life Sciences and Biomedicine, Westlake University
Sun Yi, Westlake Laboratory of Life Sciences and Biomedicine, Westlake University
Reinhard Köster, Division of Cellular and Molecular Neurobiology, Zoological Institute, Technische Universität Braunschweig
Fedor Subach, National Research Center “Kurchatov Institute”
Kiryl Piatkevich, Westlake Laboratory of Life Sciences and Biomedicine
Edward S. Boyden, Massachusetts Institute of Technology
In vivo imaging of model organisms is heavily reliant on fluorescent proteins with high brightness. Here we describe a practical method for rapid optimization of fluorescent proteins via directed molecular evolution in mammalian cells. We employed this approach to enhance intracellular brightness of a set of the spectrally diverse fluorescent proteins. The developed proteins demonstrated high performance for fluorescent labeling of neurons in culture and in vivo in model organisms such as C.elegans, Drosophila fruit flies, and zebrafish. The described method has great potential to be adopted by protein engineers due to its simplicity and convenience. We also believe that the new enhanced fluorescent proteins will find wide application for in vivo imaging of small model organisms.