BODIPY-BASED CELL PERMEABLE BACKGROUND-FREE FLUORESCENT COMPOUNDS FOR LIVE CELL IMAGING

Abstract

LABELING A SPECIFIC BIOMOLECULE IN NATIVE LIVE CELL, PARTICULARLY WITH FLUORESCENT PROBES, IS A POWERFUL STRATEGY TO ACHIEVE IN-DEPTH UNDERSTANDING OF ITS BIOLOGICAL ROLES AND FUNCTIONS. BESIDES NONTOXIC AND SPECIFIC, THE DESIRABLE PROBES FOR INTRACELLULAR LABELING SHOULD BE HIGHLY CELL PERMEABLE WITHOUT NONSPECIFIC INTERACTIONS WITH OTHER CELLULAR COMPONENTS. TO DATE, MOST REPORTED PROBES, EITHER DO NOT CROSS CELL MEMBRANE OR LEAVE NONSPECIFIC BACKGROUND, RESULTING IN LIMITED APPLICABILITY FOR LIVE CELL LABELLING. WE REPORT A PREDICITIVE MODEL TO GUIDE THE DEVELOPMENT OF BACKGROUND-FREE FLUORESCENT PROBE VIA HIGH THROUGHPUT PHENOTYPIC SCREENING IN COMBINATION WITH CHEMINFORMATICS. USING THE MODEL AS A GUIDE, WE DEVELOPED BO-1 AND BO-3, PHOTOSTABLE CYCLOOCTYNE-CONTAINING BODIPY PROBES, TO SPECIFICALLY LABEL VARIOUS INTRACELLULAR ORGANELLES AND ENGINEERED PROTEINS IN LIVE CELLS WITH MINIMUM BACKGROUND. SUBSEQUENTLY, WE DEVELOPED TAMING-CF3, ANOTHER BACKGROUND-FREE PROBE THAT CAN BIND S