SynBac: Synthetic Baculovirus Genome for Next-generation Drug Discovery

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Baculovirus is a highly efficient delivery system for recombinant genes into eukaryotic cells, with great impact on the production of eukaryotic proteins, including high-value drug targets. Vaccines against cervicular cancer and others are produced by this method. More recently, baculovirus has emerged as a versatile tool for gene therapy. We contributed to the field the award-winning MultiBac technology for multiprotein complex research. These applications, at the forefront of modern biology, rely on a large baculovirus genome (130 kb) derived from AcMNPV. This genome has been intensively researched, mainly by entymologists. Genes essential for propagation in nature and in cell culture were delineated and DNA elements which impede applications in the laboratory. Genetic alterations of the wild-type viral genome have been performed, by classical knock-out technologies, to improve gene insertion, delivery and protein production properties. Such alterations require excessive effort by specialists. Therefore, it is currently not possible to fully exploit the vast potential of the baculovirus system. In the present project, we boldly propose to fully reverse the current approach. We will design in silico and construct in vitro new, fully synthetic customized baculovirus genomes which will be, for the first time, in a streamlined, highly versatile format for multigene transfer and the production of high-value, next generation recombinant protein targets for drug discovery. We will apply state-of-the-art genome editing tools, notably CRISPR-Cas9, to inform our approach by systematically disrupting genes and non-coding regions including gene regulatory elements. We further aim to address the "scale-up problem" which currently impedes pharma-scale biologics production by baculoviral systems. As proof-of-concept, we already created a partly synthetic hybrid genome by replacing a large part (20 kb) of wild-type with designer DNA. Rigorous testing of this prototype compellingly validated our approach.The CASE fellow will:(1) utilize computational biology, comparative bioinformatics and data mining to create blueprints for optimized minimal baculovirus genomes. (2) Synthesize designer genomes in large fragments, and use advanced recombination technologies to assemble these into functional genomes (AstraZeneca platform).(3) Exploit cutting-edge CRISPR-Cas9 tool to edit genes and regulatory DNA elements in the baculoviral genome in a parallelized fashion (AstraZeneca platform) and implement this information in the synthetic design. (4) Address the "scale-up problem" by reconfiguring the very late phase of baculoviral life cycle ("hyperburst management"). (5) Rigorously validate novel designer genomes experimentally.These will be the first fully synthetic baculoviral genomes, with the potential to transform academic and industrial R&D applications.