Professor
Kristian Muller
Department of Cellular and Molecular Biotechnology, Faculty of Technology, Bielefeld University, Germany
Title of the talk: Adeno-associated Virus (AAV) Based DNA and Protein Delivery Strategies
Abstract
AAVs are the preferred vector choice for gene replacement therapy with seven approved drugs in the USA and Europe to date. However, high manufacturing cost and low but present immunogenicity and heterogeneity limit their use. Approved, recombinant AAVs consist of a protein capsid enclosing a gene of interest flanked by inverted terminal repeats (ITRs). Deconstructing or modifying such AAVs allows us to leverage existing knowledge while elucidating new modes of production and application. We deployed a rolling circle amplification strategy to generate rAAV genomes and demonstrated their utility for rAAV production in HEK-293 cells as well as for packaging within lipid-nanoparticles. These nanoparticles were co-delivered with mRNA coding the AAV Rep integrase and we found genome integration similar to that of wtAAV. AAV capsid proteins VP3 were expressed in E. coli and functional particles were obtained by refolding or directly in the cytosol. We also endowed capsid proteins via terminal fusion or loop insertion with additional functional domains providing binding, reporting or antigen presentation. This modular approach to AAV components enhances rAAV production and paves the way for new, widely applicable therapies and vaccines.
Biography
Kristian Müller is a professor of cellular and molecular biotechnology at Bielefeld University, Germany. He was an assistant professor and group leader at the University of Freiburg and the University of Potsdam. Prior to his independent career, he worked as a postdoctoral scientist at the University of California at Berkeley, and he received his Ph.D. from the University of Zurich and his Diploma in Biochemistry from the University of Hannover. He combines gene with protein engineering in the context of synthetic biology and cellular systems to drive basic research and create innovative biotherapeutics
Publications: Researchgate, Google Scholar, ORCID.