421-Takenouchi-AgedYoungOocyte/Fig1B_Chr11Probe
Author reviewed
Z-series images of mouse oocytes visualizing H2B-SNAP and chromosome11
Dataset ID
Fig1B_Chr11Probe
Title
Z-series images of mouse oocytes visualizing H2B-SNAP and chromosome11
Description
Z-series images of mouse oocytes visualizing H2B-SNAP and chromosome1. For CRISPR-Sirius method, single-guide RNAs (sgRNAs) to detect individual chromosome-specific repeat region were prepared. The repeat sequences of target chromosomes are labeled by noncleaving Cas9 (dCas9), sgRNA with aptamers,
and fluorescent protein–fused aptamer-binding proteins.
channel1; probe for chromosome11, channel2; H2B-SNAP, channel3; bright field
Folder Name
Fig1B_Chr11Probe
Biosample
Description
Mouse oocytes at 5 hours after GVBD (germinal vesicle breakdown), in which H2B-SNAP and chromosome11 were visualized.
Extrinsic Variable
To visualize chromosome, in vitro transcribed mRNAs for H2B-SNAP was injected.To label the chromosome11,
mRNA for PCP-3mClover, dCas9 protein, and sgRNA for chromosome1 were injected.
Ontology
Genetics
Protein / Probe
-single-guide RNA
targeting repeat sequences of the chromosome11
Method / Genotype
injection
Protein / Probe
Histone H2B type 1-J
Method / Genotype
injection
Treatments
Imaging Method
Detection Method
photomultiplier tube (PMT)
Visualization Method
H2B with SNAP-tag was visuallised with SNAP-Cell 647-SiR. The specifc chromosome was visualized with mClover.
Illumination Method
single point scanning
Source of Contrast
distribution of a specific nucleic acid sequence
Imaged Parameter
fluorescence emission
Sample Preparation Method
living tissue
Instrument
Objective
Zeiss 40×C-Apochromat 1.2NA