Dataset ExtFig6a_chromosome_fast
Project
387-Ando-StayGold
Title
Super resolution cellular images using fluorescent-tags of StayGold variants
Description
Although StayGold is a bright and highly photostable fluorescent protein, its propensity for obligate dimer formation may hinder applications in molecular fusion and membrane targeting. To attain monovalent as well as bright and photostable labeling, the authors engineered tandem dimers of StayGold to promote dispersibility. On the basis of the crystal structure of this fluorescent protein, they disrupted the dimerization to generate a monomeric variant that offers improved photostability and brightness compared to StayGold. The authors applied the new monovalent StayGold tools to live-cell imaging experiments using spinning-disk laser-scanning confocal microscopy or structured illumination microscopy. They achieved cell-wide, high-spatiotemporal resolution and sustained imaging of dynamic subcellular events, including the targeting of endogenous condensin I
to mitotic chromosomes, the movement of the Golgi apparatus and its membranous derivatives along microtubule networks, the distribution of cortical filamentous actin and the remolding of cristae membranes within mobile mitochondria.
Funding
This work was supported in part by Grant-in-Aid for Scientific Research (S) (21H05041 to A.M.), Grant-in-Aid for Innovative Areas: Resonance Bio (15H05948 to A.M.) and Information Physics of
Living Matters (19H05794 and 19H05795 to Y.O.), Japan Science and Technology Agency Core Research for Evolutionary Science and Technology program, ‘Spatiotemporal dynamics of intracellular components’ (JPM JCR20E2 to Y.O.) and Moonshot R&D program ‘Realization of ultra-early disease prediction and intervention by
2050’ (JPM JMS2025-14 to Y.O.), Marine Biomass Innovation Project (NFRFT-2020-00452 to A.M.), the Brain Mapping by Integrated Neurotechnologies for Disease Studies from AMED (Brain/MINDS, JP15dm0207001 to A.M.), RIKEN Collaboration Seed Fund (to M.T. and R.A.) and Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)) from AMED (JP21am0101070 to M.Y.).
Title
Time-lapse images of CAP-H-td5oxStayGold and SiR-DNA-labeled chromosomes in HCT116 cells
Description
Time-lapse images of CAP-H-td5oxStayGold and SiR-DNA-labeled chromosomes in HCT116 cells. With CRISPR-Cas9 genome-editing technique, a subunit of condensin I (CAP-H) was endogenously tagged at the C terminus with td5oxStayGold (CAP-H-td5oxStayGold) in the HCT116 cells.
Description
Genome-edited HCT116 cells in which a subunit of condensin I (CAP-H) was endogenously tagged at the C terminus with td5oxStayGold
Intrinsic variables
HCT116 cells
were edited by CRISPR-Cas9 genome-editing technique, in which a subunit of condensin I (CAP-H) was endogenously tagged at
the C terminus with td5oxStayGold
Gene ontology: Biological processes
Gene ontology: Cellular components
Gene ontology: Molecular functions
Protein names
Condensin complex subunit 2 (
Q15003)
Protein tags
td5oxStayGold
Dimensions
1024x1024x1x2x1801,
512x512x1x3x1
Microscopy types
spinning disk confocal microscopy (
FBbi:00000253)
(confocal microscopy (LSCM))
Visualization methods
probes for nucleic acid
td5oxStayGold, a StayGold derivative
Contrast enhancing methods
Resolution enhancing methods
Sample preparation methods
living tissue
Body
Olympus-Evident IX83
Detector
Hamamatsu ORCA-Flash4.0 CL, USB
Objective
Olympus-Evident UPLAPO100XOHR