Enhancing rAAV gene therapy manufacturing: Improving rAAV productivity & genome packaging through plasmid and cell engineering

The use of gene therapy products for the treatment of a large range of indications and as potential vaccines in the clinic shows great promise but producing high titre viral vectors and manufacturing at scale remains a challenge and contributes to the cost of these therapies.
At the University of Kent we have used HET293 cells to produce viral vectors, particularly adenovirus associated virus (AAV), for the delivery of gene therapies and through our studies and the literature identified cellular targets that are increased or decreased in expression when comparing conditions leading to lower or amounts of correctly assembled and packaged AAVs. The proposed PhD project will use a combination of genome editing, siRNA knockdown and over-expression studies to determine the impact of manipulation of individual and multiple targets on AAV production from HEK293 cells, thus furthering our understanding of the basic cell biology that underpins AAV production from HEK293 cells and creating novel, industrially relevant HEK293 cell lines for enhanced AAV production.
Impact: The project has the potential to deliver a step-change in our understanding of the fundamental biology underpinning the responses of HEK293 cells to an imposed rAAV load and how these define the efficiency of vector packaging, alongside application of this knowledge to generate new HEK293 hosts for improved manufacturing of rAAV vectors at vastly reduced cost.