Time-course relative transcriptional profiles of the targets controlled by the artificial control system (hQSCS) in its ON and OFF states.

To confirm that hQSCS can simultaneously regulate multiple functional modules and maintain differential expression compared to the OFF state over an extended period, we performed time‑course transcriptome analysis on the metabolically engineered industrial strain N9 at multiple time points.

Specifically, hQSCS was kept in the OFF state by adding cellulose as an inducer to serve as the control group, whereas the experimental group naturally grew and autonomously triggered and maintained hQSCS in the ON state during the stationary phase. Samples were collected at different growth stages for transcriptome sequencing. Differential expression analysis of this dataset will help determine whether hQSCS can effectively control a metabolic network orthogonal to the endogenous system, which is crucial for the broader application of hQSCS and provides insights for the implementation of orthogonal control systems in other rare actinomycetes.