Single-Molecule Nanoscopy Elucidates RNA Polymerase II Transcription at Single Genes in Live Cells

6Citations
Citations of this article
195Readers
Mendeley users who have this article in their library.

Abstract

Transforming the vast knowledge from genetics, biochemistry, and structural biology into detailed molecular descriptions of biological processes inside cells remains a major challenge—one in sore need of better imaging technologies. For example, transcription involves the complex interplay between RNA polymerase II (Pol II), regulatory factors (RFs), and chromatin, but visualizing these dynamic molecular transactions in their native intracellular milieu remains elusive. Here, we zoom into single tagged genes using nanoscopy techniques, including an active target-locking, ultra-sensitive system that enables single-molecule detection in addressable sub-diffraction volumes, within crowded intracellular environments. We image, track, and quantify Pol II with single-molecule resolution, unveiling its dynamics during the transcription cycle. Further probing multiple functionally linked events—RF-chromatin interactions, Pol II dynamics, and nascent transcription kinetics—reveals detailed operational parameters of gene-regulatory mechanisms hitherto-unseen in vivo. Our approach sets the stage for single-molecule studies of complex molecular processes in live cells. Key limitations of imaging single copies of highly abundant factors at specific intracellular sites of activity are overcome by an active target-locking system that achieves long-term tracking at single-molecule resolution.

Cite

CITATION STYLE

APA

Li, J., Dong, A., Saydaminova, K., Chang, H., Wang, G., Ochiai, H., … Pertsinidis, A. (2019). Single-Molecule Nanoscopy Elucidates RNA Polymerase II Transcription at Single Genes in Live Cells. Cell, 178(2), 491-506.e28. https://doi.org/10.1016/j.cell.2019.05.029

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free