GSE108442
Genome Engineering for Enhancing Recombinant Protein Expression in E.coli
Overall aim of this study was to investigate the role of up-regulated genes  in the generation of cellular stress response that is triggered upon induction of recombinant protein synthesis. Up-regulated genes identified previously by transcriptomic analysis,  were knocked out from the host genome  and their impact on cellular health and  expression capabilities was analyzed. Transcriptomic profiling of  the top performing  double knock out (?(elaA+cysW)  having significantly higher  protein expression levels was compared with the control to demonstrate the ability of this knock-out strain to counter cellular stress. Overall design: Total RNA obtained was isolated from control (E.coli BW25113), single knock-out strains i.e. ?elaA & ?cysW & the combined  double knock-out (?elaA+?cysW) . Samples were collected at two time-points i.e. pre-induction(0h) & post-induction(6h). Total  9 samples  were sent to AgriGenome for library preparation & RNA sequencing.