10073149UPEC SimCell as a potential vaccine against UTIClosedCollaborative R&DInnovate UKUsing live bacteria as vaccines and therapies for diseases has long been seen as an attractive idea, offering a way to target the immune system more comprehensively. Unfortunately, the approach is difficult to control, with potentially dangerous side-effects. SimCell technology, developed at the University of Oxford, solves this problem by producing bacterial cells that lack genetic material (DNA) and are therefore unable to divide. SimCells are made by introducing an engineered switch that can be triggered to destroy the bacteria's DNA so that it can no longer divide. The bacteria remain intact, retaining the important cell-surface features recognised by the immune system. This contrasts with existing methods of inactivating bacteria using heat, chemicals, or irradiation are harsh treatments that damage the cells and reduce their ability to induce immune responses. In this project, we will adopt synthetic biological methods to engineer uro-pathogenic Escherichia coli (UPEC) into SimCells. The key goal is to achieve tight regulation of the endonuclease expression system to ensure maximum efficiency of SimCell conversion and prevent the emergence of escape cells. This includes the use of rationally designed promoter systems combined with the application of a robotic platform to optimize the expression and working conditions of the endonuclease upon induction. A successful project outcome will help us select a lead UTI SimCell vaccine candidate to enter the lead optimization stage and accelerate the development of other SimCell vaccine pipelines of Gram-negative bacteria. Ultimately, this project will result in the engineering of a bacterial vaccine prototype that could not only help reduce the inappropriate use of antibiotics but also positively impact the health of 50% of all women in the world.