Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping

Abstract

The recombinant baculoviruses were constructed to investigate the necessity of VSV-G pseudotyping for mammalian cell transduction. The viruses were designed to express green fluorescent protein (GFP) gene under the control of cytomegalovirus promoter, with or without pseudotyping with VSV-G. VSV-G was placed under the control of polyhedrin promoter that is recognized by insect cells, allowing the formation of pseudotyped baculovirus. The study findings demonstrate that the pseudotyping of baculovirus significantly enhanced transduction efficiency compared to non-pseudotyped baculovirus, resulting in consequent distinction in the expression of GFP in mammalian cells. The results confirmed that pseudotyping is important for baculovirus mediated gene delivery. Further, when full-length VSV-G pseudotyping was compared with truncated VSV-G containing GED domain (G-stem of ectodomain in conjunction with the TM and CT domains of the glycoprotein), latter was relatively less efficient in transducing mammalian cells. This study demonstrated that pseudotyping with full-length VSV-G had better transduction efficiency in mammalian cells. However, at higher multiplicity of infection, both fulllength and truncated VSV-G showed equivalent transduction. This study established the significance of pseudotyping of baculovirus with full-length VSV-G for efficient transduction of mammalian cells, utilizing the highly sensitive GFP marker system. These findings have significant implications in designing of baculovirus vector based antigen delivery for developing new generation vaccines.

Solutions against emerging infectious and noninfectious human diseases through the application of baculovirus technologies

“…Thus, several investigations have been reported that have offered improvements in baculoviral vectors for BacMamtype applications. In this, list of developments can be mentioned: (i) the pseudo-typed BVs, which constructs express in insect cells the stomatitis-vesicular-virus glycoprotein (Barsoum et al 1997;Kolangath et al 2014) or other factors from eukaryote parasites and viruses (Tamura et al 2016;Hu et al 2019a) to increase transduction capability in mammals; (ii) the development of baculoviral-hybrid vectors that achieve a longer duration of the GOI expression by, for example, the presence of the ORI P of the Epstein-Barr virus, either alone (Shan et al 2006;Suzuki et al 2009) or combined with the FLP/FRP, ɸC31/attB-P or Cre/Loxp recombinase systems (Lo et al 2009(Lo et al , 2017Sung et al 2013); (iii) the inclusion of the inverted terminal repeats and the Rep gene of the adeno-associated virus (Wang 2008) or the inverted terminal repeats and the transposase gene of the Sleeping-Beauty transposon et al 2014); and (iv) genome modifications to increase the BV yield in insect cells (Graves et al 2018), among other optimizations. In addition, progress has been made in different formulations to achieve better results after the administration of recBVs in mammals.…”

Improving Baculovirus Transduction of Mammalian Cells by Incorporation of Thogotovirus Glycoproteins

“…ThGP and DhGP improved the baculovirus transduction efficiency from 4 to 12 fold compared to Ac-WT in six of the 12 selected cell lines we tested at an MOI of 5 and showed tropism of wide range of cell types, including the epithelial cells (A549, HeLa and Hep2), muscle cells (RD), bone marrow neuroblast cells (SH-SY5Y) and adrenal adenocarcinoma cells (SW13). VSV G has been widely used to enhance baculovirus transduction efficiency and expand the cell tropism (Barsoum et al 1997;Kolangath et al 2014). Here, we found that the transduction efficiency of Ac-VSVG is higher than Ac-WT in nine cell lines, but increased by more than four-fold only in three of them at an MOI of 5.…”

“…An alternative method is to incorporate heterologous GPs from viruses that infect mammalian cells, such as influenza virus neuraminidase, vesicular stomatitis virus G protein (VSV G) and the GP of rabies virus (Barsoum et al 1997;Borg et al 2004;Tani et al 2003). Among these, displaying VSV G is the most widely adopted, which can enhance the transduction of AcMNPV up to 15 fold (Kolangath et al 2014). However, VSV G exhibits significant cytotoxicity in the expressed cells and results in lower baculovirus production (Kaikkonen et al 2006).…”

Targeting Extracellular Vesicles to Dendritic Cells and Macrophages

“…In our study, we enhanced the specificity of targeted delivery by using a truncated VSV-G. This VSV-G variant comprises only the core part of the protein [ 24 ], which is responsible for the budding of extracellular vesicles from the producer cell and the release of the vesicle contents inside the target cell. In this case, it is possible to use a truncated VSV-G sequence combined with a recombinant nanobody capable of highly specific interaction with the target cell, without losing the functionality of the resulting extracellular vesicles.…”