Gene expression is a complex and highly regulated process involving numerous steps, and is challenging to explore within living cells. One of the most important steps is called transcription. It results in an mRNA strand that exits the nucleus and then translates into a protein. Nevertheless, the way that transcription functions in time and space are not yet fully understood.
In collaboration with Dr. Yaron Shav-Tal’s lab, we developed a unique system to observe a single copy gene transcription integrated into the DNA of a human cell. With this system we can investigate transcription on a single-copy gene level in living cells. By probing fluorescence fluctuations at transcription sites we extract important parameters on the dynamics controlling transcription.
Using live cell imaging techniques we study the dynamics of transcription: transcription rates and variation in the number of RNA polymerases transcribing on the gene. Studying this process on an almost single molecule level enables us to quantify a single event statistics without the obscurants of an average on multiple genes.
- S. Yunger, L. Rosenfeld, Y. Garini and Y. Shav-Tal, Single-allele analysis of transcription kinetics in living mammalian cells, Nature methods 7, 631-633 (2010).
- S. Yunger, L. Rosenfeld, Y. Garini and Y. Shav-Tal, Quantifying the transcriptional output of single alleles in single living mammalian cells, Nature Protocols, 8, 393–408 (2013).