Gene expression is a complex and highly regulated process involving numerous steps, and is challenging to explore within living cells. While in previous mammalian systems tandem gene-arrays were utilized to probe transcription site kinetics, such gene-arrays could not portray the true situation of single-alleles genomic structure. We used a system for in vivo visualization and analysis of transcriptional kinetics of single gene copy in real time [1]. We obtained high-resolution transcription measurements of a single gene under different conditions. Our work included development of a dynamic model that allows to extract important parameters of transcription combined with fluorescence recovery after photobleaching (FRAP) measurements.

[1] Yunger, S., L. Rosenfeld, Y. Garini, and Y. Shav-Tal, Single-allele analysis of transcription kinetics in living mammalian cells. Nature Methods 7, 631-633 (2010).