Transcription is the first step in the central dogma of molecular biology, when genetic information encoded on DNA is made into RNA. How this fundamental process occurs in eukaryotes is well characterized in vitro, but surprisingly poorly understood at the cellular level. This is in part because conventional live cell imaging approaches fail to capture with sufficient quantitative detail the intricate, weak and transient molecular interactions that regulate eukaryotic transcription process in vivo. To surmount these limitations, we propose novel quantitative imaging methods that enable probing eukaryotic transcription with high spatial and temporal resolution, with single molecule sensitivity directly n live mammalian cells. We investigate how spatiotemporal organization and dynamics of RNA Polymerase II (Pol II), the molecular enzyme responsible for the transcription of all protein encoding genes, regulate gene expression in individual living cells. We examine how foci of clustered Pol II, hypothesized to be foyer of eukaryotic transcriptional activity, dynamically correlate with nascent messenger RNA output at an active gene locus in the living cell. We envision determining quantitatively with single molecule sensitivity, the mechanisms by which key steps in eukaryotic transcription are dynamically regulated through the recruitment of individual molecular partners in living cells.

Public Health Relevance

Improved understanding of eukaryotic transcription processes in the cellular context will have broad implications for human health: the regulation of transcription is primordial for cellular homeostasis, and aberrant cellular regulation of transcription has been linked to many maladies including cancer and developmental disabilities.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
NIH Director’s New Innovator Awards (DP2)
Project #
1DP2CA195769-01
Application #
8755831
Study Section
Special Emphasis Panel (ZRG1-MOSS-C (56))
Program Officer
Mietz, Judy
Project Start
2014-09-23
Project End
2019-08-31
Budget Start
2014-09-23
Budget End
2019-08-31
Support Year
1
Fiscal Year
2014
Total Cost
$2,340,000
Indirect Cost
$840,000
Name
Massachusetts Institute of Technology
Department
Physics
Type
Schools of Arts and Sciences
DUNS #
001425594
City
Cambridge
State
MA
Country
United States
Zip Code
02142