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.