Detail of NMP-R_dCas9_MS2
Project
Title
Time-lapse fluorescence microscopy images of mouse embryonic stem cells (NMP-R mESCs) transfected with the MS2 sigRNA expression vector of nuclease-dead Cas9 fused to the green fluorescent protein (dCas9-GFP) and MS2 coat protein fused to the tandem near-infrared red fluorescent protein (MCP-tdiRFP) signals
Description
NA
Release, Updated
2016-10-03,
2018-11-15
2018-11-15
License
CC BY-NC
Kind
Image data
based on Experiment
File Formats
Data size
4.6 MB
Protein names
dCas9, MCP
Protein tags
GFP, tdiRFP
Datatype
gene expression dynamics
Molecular Function (MF)
Biological Process (BP)
cellular protein localization
(
GO:0034613
)
Cellular Component (CC)
-
Biological Imaging Method
XYZ Scale
XY: 0.16 micrometer/pixel, Z: 0. micrometer/slice
T scale
2 minute for each time interval
Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-
Summary of Methods
See details in Ochiai et al. (2015) Nucleic Acids Research, 43(19): e127
Related paper(s)
Hiroshi Ochiai, Takeshi Sugawara, Takashi Yamamoto (2015) Simultaneous live imaging of the transcription and nuclear position of specific genes., Nucleic acids research, Volume 43, Number 19, pp. e127
Published in 2015 Oct 30 (Electronic publication in June 19, 2015, midnight )
- doi: 10.1093/nar/gkv624
-
pmid: 26092696
(Abstract) The relationship between genome organization and gene expression has recently been established. However, the relationships between spatial organization, dynamics, and transcriptional regulation of the genome remain unknown. In this study, we developed a live-imaging method for simultaneous measurements of the transcriptional activity and nuclear position of endogenous genes, which we termed the 'Real-time Observation of Localization and EXpression (ROLEX)' system. We demonstrated that ROLEX is highly specific and does not affect the expression level of the target gene. ROLEX enabled detection of sub-genome-wide mobility changes that depended on the state of Nanog transactivation in embryonic stem cells. We believe that the ROLEX system will become a powerful tool for exploring the relationship between transcription and nuclear dynamics in living cells.(MeSH Terms)
- Animals
- Capsid Proteins/genetics
- Cell Nucleus/genetics[major]
- Cells, Cultured
- Embryonic Stem Cells/metabolism
- Endonucleases/analysis
- Endonucleases/genetics
- Gene Expression
- Green Fluorescent Proteins/analysis
- Green Fluorescent Proteins/genetics
- Homeodomain Proteins/genetics
- In Situ Hybridization, Fluorescence[major]
- Mice
- Mice, Inbred C57BL
- Nanog Homeobox Protein
- Optical Imaging/methods
- Pluripotent Stem Cells/metabolism
- Recombinant Fusion Proteins/analysis
- Transcription, Genetic[major]
Contact
Hiroshi Ochiai
, Hiroshima University
, Research Center for the Mathematics on Chromatin Live Dynamics (RcMcD)
Contributors
Hiroshi Ochiai, Takeshi Sugiwara, Takashi Yamamoto
OMERO Dataset
OMERO Project
External Link