A Baculovirus Expression Vector Derived Entirely from Non-Templated, Chemically Synthesized DNA Parts

Baculovirus expression system1s are a widely used tool in recombinant protein and biologics production. To enable the possibility of genome modifications unconstrained through low-throughput bespoke classical manipulation techniques, we set out to construct baculovirus vector (>130 kb dsDNA) built from modular, chemically synthesized DNA parts. We constructed synthetic version Autographa californica multiple nucleopolyhedrovirus (AcMNPV) two steps hierarchical Golden Gate assembly. Over 140 restriction endonuclease sites were removed discrimination native genomes. A head-to-head comparison our AcMNPV with vectors showed no significant difference growth kinetics or adeno-associated virus production—suggesting that neither replication nor very-late gene compromised by design assembly method. With unprecedented control over at single-nucleotide level, hope ambitiously explore novel streamlined for production development.