Production and delivery of active therapeutic proteins into their target cells for treatment of disease is still an important unsolved challenge in the biopharmaceutical sector. We have developed an innovative recombinant E.coli-based expression system that induces packaging of a range of recombinant proteins in membrane bound vesicles, thus enabling production of otherwise challenging insoluble, toxic, disulphide bond containing recombinant proteins in bacteria. Preliminary studies with these vesicles show that they can fuse with human cells in culture, thus delivering their cargo into the cell. The aim now is to modify the surface of these vesicles so that we can selectively target them to particular disease cell types. Approach: The proof of concept will be to express the PD-1 protein, which binds to PD-L1 found overexpressed on the surface of multiple cancers, on these vesicles (Kent) with a recombinant GFP protein as the vesicle cargo. These vesicles will then be isolated and incubated with different cancer cell types that do/do not express PD-L1 to evaluate cellular selectivity (Kent) and analyse gene expression changes by RNA sequencing (Southampton). Areas of potential impact: The potential impact of this project is that it will deliver proof-of-concept that an active protein can be recombinantly expressed in E.coli generated vesicles and selectively targeted to enter a specific human cell type. This is a technology that has the potential to be applied to a wide range of biotechnology and medical applications, including the diagnosis and treatment of multiple diseases including Parkinson's, Alzheimer's and cancer. In this project the student will gain experience in a wide range of cell biological, molecular, biophysical and biotechnological techniques, working with researchers and academics with a wealth of experience in these areas.