RT Journal Article
SR Electronic
T1 Computer Designed PRC2 Inhibitor, EBdCas9, Reveals Functional TATA boxes in Distal Promoter Regions
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.11.16.385922
DO 10.1101/2020.11.16.385922
A1 Shiri Levy
A1 Logesh Somasundaram
A1 Infencia Xavier Raj
A1 Diego Ic-Mex
A1 Sven Schmidt
A1 Ammar Alghadeer
A1 Henrik Honkanen
A1 R. David Hawkins
A1 Julie Mathieu
A1 Yuliang Wang
A1 David Baker
A1 Karol Bomsztyk
A1 Hannele Ruohola-Baker
YR 2020
UL http://biorxiv.org/content/early/2020/11/17/2020.11.16.385922.abstract
AB 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 a single nucleosome 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 resolution 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 is broadly applicable for epigenetic regulation at a single locus to control gene expression.Competing Interest StatementThe authors have declared no competing interest.