Direct observation of frequency modulated transcription in single cells using light activation.
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. DOI:http://dx.doi.org/10.7554/eLife.00750.001.
Laboratory of Receptor Biology and Gene Expression , Center for Cancer Research, National Cancer Institute, National Institutes of Health , Bethesda , United States.
Daniel R
DR
Larson
Christoph
C
Fritzsch
Liang
L
Sun
Xiuhau
X
Meng
David S
DS
Lawrence
Robert H
RH
Singer
D006801:Humans; D008027:Light; D012333:RNA, Messenger / Q000096:biosynthesis; D014158:Transcription, Genetic
D012333:RNA, Messenger
Human; fluorescence; gene expression; single-molecule; transcription factors