GEO Logo
   NCBI > GEO > Accession DisplayHelp Not logged in | LoginHelp
GEO help: Mouse over screen elements for information.
Series GSE195552 Query DataSets for GSE195552
Status Public on Feb 12, 2022
Title dCas9 Fusion to Computer Designed PRC2 Inhibitor Reveals Functional TATA Box in Distal Promoter Region [ChIP-seq]
Organism Homo sapiens
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary The critical process in development, bifurcation of cellular fates, requires epigenetic H3K27me3 marks propagated by PRC2 complex. However, the precise chromatin loci of functional H3K27me3 marks are not yet known. Here we identify critical PRC2 functional sites at high resolution. We fused a computationally designed protein, EED binder (EB) that competes with EZH2 and thereby disrupts PRC2 function, to dCas9 (EBdCas9) to direct PRC2 inhibition at a precise locus using gRNA. We targeted EBdCas9 to 4 different genes ( TBX18, p16, CDX2 and GATA3 ) and observed epigenetic remodeling at a single nucleosome resulting in gene activation. Remarkably, while traditional TATA box is located 30bp upstream of TSS, we identified a functional TATA box, >500bp of TSS, normally repressed by PRC2 complex. Deletion of this TATA box eliminates EBdCas9 dependent TBP recruitment and transcriptional activation. Targeting EBdCas9 to CDX2 and GATA3 results in trophoblast trans-differentiation. EBdCas9 technology may provide a broadly applicable tool for epigenetic regulation at a single locus to control gene expression
Overall design biological samples were isolated from iPSC WTC cell lines expressing either EB-Flag or EBdCas9 for 3 days, or no expressing cells. ChIP-seq H3K27me3 was preform for each sample.
Contributor(s) Ng WI, Ruohola-Baker H
Citation(s) 35235780
Submission date Jan 27, 2022
Last update date Apr 19, 2022
Contact name Hannele Ruohola-Baker
Organization name Institute for Stem Cell and Regenerative Medicine
Street address 850 Republican Street
City Seattle
State/province WA
ZIP/Postal code 98109
Country USA
Platforms (1)
GPL30173 NextSeq 2000 (Homo sapiens)
Samples (4)
GSM5840471 EBdCas9+Dox
GSM5840472 EBdCas9-Dox
GSM5840473 EB-Flag+Dox
This SubSeries is part of SuperSeries:
GSE195555 dCas9 Fusion to Computer Designed PRC2 Inhibitor Reveals Functional TATA Box in Distal Promoter Region
BioProject PRJNA801245

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE195552_RAW.tar 565.2 Mb (http)(custom) TAR (of BW)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

| NLM | NIH | GEO Help | Disclaimer | Accessibility |
NCBI Home NCBI Search NCBI SiteMap