跳转至内容
Merck
  • Single-cell profiling reveals that eRNA accumulation at enhancer-promoter loops is not required to sustain transcription.

Single-cell profiling reveals that eRNA accumulation at enhancer-promoter loops is not required to sustain transcription.

Nucleic acids research (2016-12-10)
Samir Rahman, Cornelia E Zorca, Tatiana Traboulsi, Emmanuel Noutahi, Matthew R Krause, Sylvie Mader, Daniel Zenklusen
摘要

Enhancers are intergenic DNA elements that regulate the transcription of target genes in response to signaling pathways by interacting with promoters over large genomic distances. Recent studies have revealed that enhancers are bi-directionally transcribed into enhancer RNAs (eRNAs). Using single-molecule fluorescence in situ hybridization (smFISH), we investigated the eRNA-mediated regulation of transcription during estrogen induction in MCF-7 cells. We demonstrate that eRNAs are localized exclusively in the nucleus and are induced with similar kinetics as target mRNAs. However, eRNAs are mostly nascent at enhancers and their steady-state levels remain lower than those of their cognate mRNAs. Surprisingly, at the single-allele level, eRNAs are rarely co-expressed with their target loci, demonstrating that active gene transcription does not require the continuous transcription of eRNAs or their accumulation at enhancers. When co-expressed, sub-diffraction distance measurements between nascent mRNA and eRNA signals reveal that co-transcription of eRNAs and mRNAs rarely occurs within closed enhancer-promoter loops. Lastly, basal eRNA transcription at enhancers, but not E2-induced transcription, is maintained upon depletion of MLL1 and ERα, suggesting some degree of chromatin accessibility prior to signal-dependent activation of transcription. Together, our findings suggest that eRNA accumulation at enhancer-promoter loops is not required to sustain target gene transcription.

材料
货号
品牌
产品描述

Sigma-Aldrich
多聚-L-赖氨酸 溶液, 0.1 % (w/v) in H2O
Sigma-Aldrich
单克隆抗 β-肌动蛋白抗体 小鼠抗, clone AC-15, ascites fluid
Sigma-Aldrich
Cy®3 Mono 5-pack, Cytiva PA23001
Sigma-Aldrich
Cy®5 Mono 5-pack, Cytiva PA25001