Report

Transcription-coupled changes in nuclear mobility of mammalian cis-regulatory elements

See allHide authors and affiliations

Science  25 Jan 2018:
eaao3136
DOI: 10.1126/science.aao3136

You are currently viewing the abstract.

View Full Text

Log in to view the full text

Log in through your institution

Log in through your institution

Abstract

To achieve guide RNA (gRNA) multiplexing and an efficient delivery of tens of distinct gRNAs into single cells, we develop a molecular assembly strategy, chimeric array of gRNA oligo (CARGO). We couple CARGO with dCas9 imaging to quantitatively measure movement of enhancers and promoters that undergo differentiation-associated activity changes in live embryonic stem cells. While all examined functional elements exhibit sub-diffusive behavior, their relative mobility increases concurrently with transcriptional activation. Furthermore, acute perturbation of Pol II activity can reverse these activity-linked increases in loci mobility. Through quantitative CARGO-dCas9 imaging, we provide direct measurements of cis-regulatory element dynamics in living cells and distinct cellular and activity states, and uncover an intrinsic connection between cis-regulatory element mobility and transcription.

View Full Text