Equipment for Spatiotemporal Dynamics of the Genome by 3D Orbital Tracking
Project Summary
Our goal is to develop 3D orbital tracking, a powerful single-molecule fluorescence fluctuation
technique, to measure the timing of gene activation, splicing, and genome organization at an active gene in
living cells. By scanning a laser in a circle around a fluorescently labeled site of transcription in a confocal
microscope, we can measure fluorescence intensity at high speed for a very long time in living cells. By
fluorescent labeling, DNA, RNA, and protein factors in different colors, we can monitor complex assembly,
transcription, splicing, and termination of RNA and the proximity of distal DNA elements like enhancers in the
living genome.
Looking at nature in a new way, or with a novel tool, often reveals previously unknown details. This proposal
describes such a project. Although much work has been done to study transcription, splicing, and termination
of RNA, they have yet to be able to characterize a complete kinetic profile of how the RNA is synthesized from
a gene and then released or when splicing occurs. With such a tool, we may see much that was previously
invisible. We can begin to discern complex biochemical reactions that can only be measured in living cells.
This research project will support interdisciplinary undergraduate, graduate, and postdoctoral trainees, who will
learn advanced techniques in genome biology and single-molecule imaging, shedding new light on the
spatiotemporal dynamics of transcription and splicing. Project members will participate in activities to broaden
the participation of underrepresented groups in science, mathematics, engineering, and technology.