RNA-seq analysis of sysRNA expression arrays for ampC regulation in Escherichia coli
Synthetic small RNA (sysRNA) arrays were designed to enable modular, multiplexed regulation of ampC expression in Escherichia coli K-12 substrain MG1655, using combinations of one to three sysRNAs (p1RybB, p2MicF, p3MicA) within a single array. RNA sequencing was performed to characterize transcriptome-wide effects of these constructs relative to a scaffold control construct lacking seed regions. Three array configurations were analyzed: p1RybB, p1RybB-p2MicF, and p1RybB-p2MicF-p3MicA. Total RNA was extracted from exponential-phase cultures (OD600 of 0.4). Strains were cultivated in LB medium at 37°C with continuous shaking (180 rpm) under aerobic conditions.
Synthetic small RNA (sysRNA) arrays were designed to enable modular, multiplexed regulation of ampC expression in Escherichia coli K-12 substrain MG1655, using combinations of one to three sysRNAs (p1RybB, p2MicF, p3MicA) within a single array. RNA sequencing was performed to characterize transcriptome-wide effects of these constructs relative to a scaffold control construct lacking seed regions. Three array configurations were analyzed: p1RybB, p1RybB-p2MicF, and p1RybB-p2MicF-p3MicA. Total RNA was extracted from exponential-phase cultures (OD600 of 0.4). Strains were cultivated in LB medium at 37°C with continuous shaking (180 rpm) under aerobic conditions.