| Popis: |
Although StayGold is a bright and highly photostable fluorescent protein (FP), its obligate dimer formation may prevent its application in molecular fusion and membrane targeting. With the objective of attaining monovalent as well as bright and photostable labeling, we engineered tandem dimers of StayGold to be dispersible. On the basis of the crystal structure of this FP, we disrupted the dimer interface to develop monomeric variants of StayGold. We applied the new StayGold tools to live cell imaging experiments using spinning-disk laser scanning confocal microscopy or structured illumination microscopy. We achieved cell-wide, high–spatiotemporal-resolution, and sustained imaging of subcellular dynamic events, including the targeting of endogenous condensin I to mitotic chromosomes at the onset of mitosis, the movement of the Golgi apparatus and its membranous derivatives along microtubule networks, the distribution of cortical filamentous actin near the plasma membrane, and the remolding of cristae membranes within mobile mitochondria. |
