Experimental strategy used for assessing the role of chromatin environment on stochastic gene expression. After generation of cellular clones expressing the fluorescent reporter mCherry, stably integrated as a unique copy into the genome, the fluorescence distributions obtained by flow cytometry ('FACS') were compared with simulated distributions generated by a two-state model ('Model'). After experimental determination and exploration of transcription-translation parameters (ρ, transcription rate; γ, translation rate; , mRNA degradation rate; , protein degradation rate and α, protein fluorescence coefficient), the best parameter sets were identified, and then used to compute the specific chromatin dynamics (kon and koff, which are, respectively, the opening and closing transition rates of the chromatin at the reporter integration site) for each clone.
Viñuelas et al. BMC Biology 2013 11:15 doi:10.1186/1741-7007-11-15