Truncated vesicular stomatitis virus G protein improves baculovirus transduction efficiency in vitro and in vivo

Kaikkonen, Minna U.; Räty, Jani K.; Airenne, Kari J.; Wirth, Thomas; Heikura, Tommi; Ylä‐Herttuala, Seppo

doi:10.1038/sj.gt.3302657

Cited by 77 publications

(82 citation statements)

References 36 publications

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“…It was earlier reported that VSV-GED had strong improvement of transduction efficiency over GP64 control [11]. To understand if the disparity between the observation in this study and the previous study [11] was due to difference in MOI used, a transduction was carried out in BHK-21 cells using the high MOI of 50 like in the previous study.…”

Section: Discussionmentioning

confidence: 65%

“…The observed variation in VSV-G mediated transduction efficiency in different cell types could be due to difference in receptor(s) expression. After confirming that VSV-G pseudotyping enhanced the transduction of mammalian cells by the baculovirus, it was investigated if VSV-GED, a truncated version of VSV-G, comprising of partial G-stem of ectodomain in juxtaposition with the TM and CT domains of VSV-G, was sufficient enough for increasing baculovirus-mediated gene delivery, as reported earlier [4,11]. Therefore the transduction was evaluated for recombinant baculovirus encoding AcGFP under CMV promoter, pseudotyped with either full-length VSV-G or its truncated version.…”

Section: Discussionmentioning

confidence: 86%

“…To understand if the disparity between the observation in this study and the previous study [11] was due to difference in MOI used, a transduction was carried out in BHK-21 cells using the high MOI of 50 like in the previous study. It was observed that at the higher MOI, both fulllength and truncated VSV-G pseudotyped virus constructs show similar transduction efficiency.…”

Section: Discussionmentioning

confidence: 93%

“…However, several researchers [9,19] have reported the utility of vesicular stomatitis virus glycoprotein (VSV-G, 511 amino acids long) pseudotyping of baculovirus for gene delivery into mammalian systems. Furthermore, while some researchers have utilized fulllength VSV-G for pseudotyping of baculovirus [9,19], there are also reports that truncated VSV-G, termed as VSV-GED, having a 21-amino-acid G-stem of ectodomain in conjunction with the transmembrane (TM) and C-terminal (CT) domain, was sufficient and increased the transduction efficiency of baculovirus-mediated gene delivery [4,11]. Thus far, there is no systematic comparative study on the identical platform investigating if VSV-G pseudotyping of the baculovirus is redundant given that the virus already has a similar functional protein, GP64.…”

Section: Introductionmentioning

confidence: 99%

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Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping

et al. 2014

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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.

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Section: Discussionmentioning

confidence: 65%

Section: Discussionmentioning

confidence: 86%

Section: Discussionmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

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Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping

et al. 2014

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“…22 Another appealing application of baculovirus is in vivo gene delivery, but most previous in vivo studies delivered only reporter genes into animal models. 23 28 show that a baculovirus expressing diphtheria toxin A inhibits the glioma xenograft growth in the rat brain, suggesting the potentials of baculovirus in cancer therapy. However, the applications of baculovirus in mesenchymal stem cell engineering and bone repair have yet to be reported.…”

Section: Discussionmentioning

confidence: 99%

Baculovirus as a new gene delivery vector for stem cell engineering and bone tissue engineering

et al. 2007

Gene Ther

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Baculovirus has emerged as a novel vector for in vitro and in vivo gene delivery due to its low cytotoxicity and nonreplication nature in mammalian cells, but the applications of baculovirus in the genetic modification of human mesenchymal stem cells (hMSCs) and tissue engineering are yet to be reported. In this study, we genetically engineered hMSCs with a baculovirus (Bac-CB) expressing bone morphogenetic protein-2 (BMP-2). Bac-CB transduction of hMSCs at a multiplicity of infection of 40 triggered effective differentiation of hMSCs into osteoblasts. Supertransduction at day 6 after initial transduction enhanced the BMP-2 expression and further accelerated the in vitro osteogenesis, as confirmed by alkaline phosphatase assay, Alizarin red staining and reverse transcription-polymerase chain reaction analysis of osteoblastic genes. Implantation of the supertransduced cells at ectopic sites in the nude mice resulted in efficient cell differentiation into osteoblasts at week 2 and induced progressive mineralization and partial bone formation at week 6, as confirmed by hematoxylin and eosin, immunohistochemical and Alizarin red staining. These data collectively demonstrated, for the first time, the potential of baculovirus in hMSCs engineering and implicated its use in bone tissue engineering.

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Pharmacological Applications of Baculovirus‐Mediated Protein Expression in Mammalian Cells

Condreay

Watson

2010

CP Pharmacology

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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.

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Truncated vesicular stomatitis virus G protein improves baculovirus transduction efficiency in vitro and in vivo

Cited by 77 publications

References 36 publications

Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping

Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping

Baculovirus as a new gene delivery vector for stem cell engineering and bone tissue engineering

Pharmacological Applications of Baculovirus‐Mediated Protein Expression in Mammalian Cells

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