MultiBac: Baculovirus-Mediated Multigene DNA Cargo Delivery in Insect and Mammalian Cells

The baculovirus/insect cell system (BICS) is widely used in academia and industry to produce eukaryotic proteins for many applications, ranging from structure analysis to drug screening and the provision of protein biologics and therapeutics. Multiprotein complexes emerged as vital catalysts of cellular function. To unlock the structure and mechanism of these essential molecular machines and decipher their function, we developed MultiBac, a BICS particularly tailored for heterologous multigene transfer and multiprotein complex production. Baculovirus is unique among common viral vectors in its capacity to accommodate very large quantities of heterologous DNA and to faithfully deliver this cargo to a host cell of choice. We exploited this beneficial feature to outfit insect cells with synthetic DNA circuitry conferring new functionality during heterologous protein expression, developing customized MultiBac baculovirus variants in the process. By altering its tropism, recombinant baculovirions can be used for highly efficient delivery of customized DNA cargo in mammalian cells and tissues. Current advances in synthetic biology greatly facilitates the construction or recombinant baculoviral genomes for gene editing and genome engineering, mediated by MultiBac baculovirus tailored to this purpose. Here, recent developments and exploits of the MultiBac system are presented and discussed.