Direct observation of frequency modulated transcription in single cells using light activation

eLife

Xiuhau Meng¹  David S Lawrence²  Liang Sun³  Robert H Singer³  Christoph Fritzsch⁴  Daniel R Larson⁵ 

[1] Department of Pharmacology, University of North Carolina, Chapel Hill, United States;Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, New York, United States;Department of Chemistry, Division of Chemical Biology and Medicinal Chemistry, University of North Carolina, Chapel Hill, United States;Institute of Molecular Biology, Johannes Gutenberg University of Mainz, Mainz, Germany;Laboratory of Receptor Biology and Gene Expression, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, United States;

关键词: transcription factor;    gene expression;    single-molecule;    fluorescence;   

DOI  :  10.7554/eLife.00750

来源: DOAJ

【 摘 要 】

Single-cell analysis has revealed that transcription is dynamic and stochastic, but tools are lacking that can determine the mechanism operating at a single gene. Here we utilize single-molecule observations of RNA in fixed and living cells to develop a single-cell model of steroid-receptor mediated gene activation. We determine that steroids drive mRNA synthesis by frequency modulation of transcription. This digital behavior in single cells gives rise to the well-known analog dose response across the population. To test this model, we developed a light-activation technology to turn on a single steroid-responsive gene and follow dynamic synthesis of RNA from the activated locus.

【 授权许可】

Unknown