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Genome-wide specificity of DNA binding, gene regulation, and chromatin remodeling by TALE- and CRISPR/Cas9-based transcriptional activators.

Authors :
Polstein LR
Perez-Pinera P
Kocak DD
Vockley CM
Bledsoe P
Song L
Safi A
Crawford GE
Reddy TE
Gersbach CA
Source :
Genome research [Genome Res] 2015 Aug; Vol. 25 (8), pp. 1158-69. Date of Electronic Publication: 2015 May 29.
Publication Year :
2015

Abstract

Genome engineering technologies based on the CRISPR/Cas9 and TALE systems are enabling new approaches in science and biotechnology. However, the specificity of these tools in complex genomes and the role of chromatin structure in determining DNA binding are not well understood. We analyzed the genome-wide effects of TALE- and CRISPR-based transcriptional activators in human cells using ChIP-seq to assess DNA-binding specificity and RNA-seq to measure the specificity of perturbing the transcriptome. Additionally, DNase-seq was used to assess genome-wide chromatin remodeling that occurs as a result of their action. Our results show that these transcription factors are highly specific in both DNA binding and gene regulation and are able to open targeted regions of closed chromatin independent of gene activation. Collectively, these results underscore the potential for these technologies to make precise changes to gene expression for gene and cell therapies or fundamental studies of gene function.<br /> (© 2015 Polstein et al.; Published by Cold Spring Harbor Laboratory Press.)

Details

Language :
English
ISSN :
1549-5469
Volume :
25
Issue :
8
Database :
MEDLINE
Journal :
Genome research
Publication Type :
Academic Journal
Accession number :
26025803
Full Text :
https://doi.org/10.1101/gr.179044.114