Pharmacological Applications of Baculovirus‐Mediated Protein Expression in Mammalian Cells

Abstract

The development of cell-based assays for cellular receptors, ion channels, and transporters requires the delivery and expression of transgenes. Viral-mediated gene delivery is a particularly attractive approach for this purpose because of its efficiency and potential to deliver genes to a wide variety of cell types. Recombinant baculoviruses, long used to deliver genes to insect cells for overexpression, also effectively transfer genes to mammalian cells. The only required modification to the virus for this purpose is the addition of transgene expression cassettes controlled by mammalian cell-active promoters. These so-called BacMam viruses are useful for developing mammalian cell-based assays for investigating the function of recombinant proteins and for assessing the action of pharmacological modulators of their function. The use of such viruses is gaining popularity because of the ease of optimizing assay conditions, the ability to deliver multiple gene products, and of their flexibility in terms of host cells and levels of transgene expression. BacMam-mediated assays may be used for studying a wide variety of target proteins and assay technologies. Described in this unit is an example of BacMam-mediated gene delivery to configure a cell-based assay for pharmacological assessment of a G protein-coupled receptor. A protocol is also provided describing the use of a GFP-expressing BacMam to assess the susceptibility of new cell lines to transduction by the virus.

Baculovirus: an Insect-derived Vector for Diverse Gene Transfer Applications

“…BacMam viruses cannot replicate in mammalian cells, providing a unique low risk biosafety profile. 13 The viruses can be used to support a variety of assay formats for different targets proteins, such as G protein coupled receptors, 125,126 transporters, 127,128 ion channels, 129,130 histones, 131 enzyme fragment complementation, 132 and others. A particularly appealing aspect of using these viruses for developing assays requiring the expression of multiple proteins is that BacMam can be titrated reproducibly according to the amount of target protein desired simply by adding more or less virus.…”

Application of Baculovirus-Insect Cell Expression System for Human Therapy

“…interferons, different receptors and neurotransmiters [3,16,17]. Apart from the biochemical studies, many of these proteins have been successfully used for diagnostics of many human disorders and different infectious agents such as bacteria, viruses and parasites in different host organisms, and also for pharmacological studies [16][17][18][19][20]. BEVS is also widely used for production of viral antigens with vaccine potential for humans and animals.…”

“…Recently BacMam viruses are also being exploited for the development of cell-based assays as tools in drug discovery and investigating the mechanism of action of many pharmacological compounds [17,20]. BacMam based assays due to the broad spectrum of target cells may be used for studying many different cellular and tissue-specific processes in response to treatment and in new drug discovery programs.…”