Orthogonal Study Of Click Reaction Pairs And Its Application In Cell Labeling

As high efficient and selective methods, click reactions are widely applied in the areas of chemistry and biology. While using single click reaction for cell imaging has been widely applied, employing multiple orthogonal click reactions tracking multiple complex intracellular system within one cell remains challenging. Our research combined two click reactions which have different reaction mechanisms between each other. One is tetrazine- alkene [4+2] inverse electron demand Diels-Alder cycloaddition, the other is CBT-Cys condensation reaction. Orthogonality between these two reactions was studied and we applied these two reactions for simultaneous labeling mitochondria and outer cell membrane of HepG2 cells. The results concluded that it is possible to use two (or n) orthogonal click reactions to track multiple organelles within one cell.