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

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Polstein, LR; Perez-Pinera, P; Kocak, DD; Vockley, CM; Bledsoe, P; Song, L; Safi, A; Crawford, GE; Reddy, TE; Gersbach, CA

Published Date

  • August 2015

Published In

Volume / Issue

  • 25 / 8

Start / End Page

  • 1158 - 1169

PubMed ID

  • 26025803

Pubmed Central ID

  • 26025803

Electronic International Standard Serial Number (EISSN)

  • 1549-5469

International Standard Serial Number (ISSN)

  • 1088-9051

Digital Object Identifier (DOI)

  • 10.1101/gr.179044.114

Language

  • eng