Cellular processes are orchestrated by a large number of biomolecules in a spatially and temporally coordinated manner within a tiny volume. To uncover the underlying organizational principles and their functional relevance, we take microscopy visualization as the primary approach to systematically map the spatial localization, temporal dynamics and activity profiles of proteins and nucleic acids. For this purpose, we have developed a fragment-tagging approach which fluorescently labels target proteins using our engineered split fluorescent proteins. This approach has enabled systematic tag knock-in in cell lines through CRISPR/Cas9-medied gene editing. Our method paves the way for the large-scale generation of endogenously tagged human cell lines for the proteome-wide analysis of protein localization and interaction networks in a native cellular context.
 

Learning Objectives: 

1. Methods to labeling endogenous proteins in cells by gene editing
2. New fluorescent label for cellular proteins