Sufficiency analysis of estrogen responsive enhancers using synthetic activators.

Autor: Ginley-Hidinger M; Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA.; Department of Bioengineering, University of Utah, Salt Lake City, UT, USA., Carleton JB; Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA.; Department of Oncological Sciences, University of Utah, Salt Lake City, UT, USA., Rodriguez AC; Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA.; Department of Oncological Sciences, University of Utah, Salt Lake City, UT, USA., Berrett KC; Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA.; Department of Oncological Sciences, University of Utah, Salt Lake City, UT, USA., Gertz J; Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA jay.gertz@hci.utah.edu.; Department of Oncological Sciences, University of Utah, Salt Lake City, UT, USA.
Jazyk: angličtina
Zdroj: Life science alliance [Life Sci Alliance] 2019 Sep 30; Vol. 2 (5). Date of Electronic Publication: 2019 Sep 30 (Print Publication: 2019).
DOI: 10.26508/lsa.201900497
Abstrakt: Multiple regulatory regions bound by the same transcription factor have been shown to simultaneously control a single gene's expression. However, it remains unclear how these regulatory regions combine to regulate transcription. Here, we test the sufficiency of promoter-distal estrogen receptor α-binding sites (ERBSs) for activating gene expression by recruiting synthetic activators in the absence of estrogens. Targeting either dCas9-VP16(10x) or dCas9-p300(core) to ERBS induces H3K27ac and activates nearby expression in a manner similar to an estrogen induction, with dCas9-VP16(10x) acting as a stronger activator. The sufficiency of individual ERBSs is highly correlated with their necessity, indicating an inherent activation potential that is associated with the binding of RNA polymerase II and several transcription factors. By targeting ERBS combinations, we found that ERBSs work independently to control gene expression when bound by synthetic activators. The sufficiency results contrast necessity assays that show synergy between these ERBSs, suggesting that synergy occurs between ERBSs in terms of activator recruitment, whereas directly recruiting activators leads to independent effects on gene expression.
(© 2019 Ginley-Hidinger et al.)
Databáze: MEDLINE