A GENETIC INTERACTION APPROACH TO DEFINE CELL ENVELOPE PATHWAYS IN ZYMOMONAS MOBILIS

The bacterial cell envelope is a key to maintaining homeostasis and a barrier to environmental stress. The Zymomonas mobilis cell envelope has been shown to play a role in tolerance to external stresses and is a promising engineering target to improve tolerance to bioenergy production conditions. However, despite its important role, the key genes and pathways required for maintaining the envelope are not fully characterized. Genetic interaction (GI) screens, in which pairs of genes are systematically perturbed, could be used to reveal gene function and pathway membership in the. Z. mobilis envelope, but no such studies have been undertaken. Here, I propose using a double knockdown approach (Double-CRISPRi) to define the genetic pathways governing the Z. mobilis cell envelope. Next, by combining Double-CRISPRi with chemical genomics, I will identify novel combinations of genetic engineering targets with diverse functions. Taken together, by elucidating cell envelope pathways and identifying combinatorial engineering targets, this work will enable rational engineering of more robust Z. mobilis strains for enhanced biofuel production.