In VivoGene Transfer in Mouse Skeletal Muscle Mediated by Baculovirus Vectors

Abstract

Baculovirus vectors are efficient tools for gene transfer into mammalian cells in vitro. However, in vivo gene delivery by systemic administration is hindered by the vector inactivation mediated by the complement system. To characterize further the gene transfer efficacy of baculovirus we examined the vector transduction efficiency in skeletal muscle. Vectors expressing vesicular stomatitis virus glycoprotein (VSV-G) in the viral envelope were generated by inserting the VSV-G coding sequence downstream of the polyhedrin promoter. Two viruses were constructed to carry either the Escherichia coli beta-galactosidase (beta-Gal) gene or the mouse erythropoietin (EPO) cDNA cloned downstream of the cytomegalovirus immediate-early promoter and enhancer. The greater gene transduction efficiency of the Bac-G-betaGal vector was confirmed by comparing the beta-Gal expression level in a variety of human and mouse cell lines with that obtained on infection with Bac-betaGal, a vector that lacks VSV-G. Similarly, a 5- to 10-fold increase in beta-Gal expression between Bac-G-betaGal and Bac-betaGal was observed when mouse myoblasts and myotubes were infected. The same increase in beta-Gal expression was detected on injection of the Bac-G-betaGal vector in the quadriceps of BALB/c and C57BL/6 mice. In contrast, a 2-fold difference in transduction was observed between these two vectors in DBA/2J mouse strain. Last, expression of EPO cDNA was detected for at least 178 days in DBA/2J mice on Bac-G-EPO injection into the quadriceps whereas EPO expression declined to normal values by 35 days postinfection in BALB/c and C57BL/6 mice. Thus, these results indicate that baculovirus may be considered a useful vector for gene transfer in mouse skeletal muscle and that persistence of expression may depend on the mouse strain used.

Baculoviruses exhibit restricted cell type specificity in rat brain: a comparison of baculovirus- and adenovirus-mediated intracerebral gene transfer in vivo

“…Previously, it has been shown that they possess the capacity to transduce a variety of mammalian cell types including mesenchymal cells, neural glial cells and hepatocytes. [1][2][3][4][14][15][16][17] Our results show that baculoviruses are able to transduce brain cells efficiently in vivo and possess a very specific cell tropism in rat brain. The choroid plexus epithelial cells were transduced with high efficiency (Figure 1, Table 1).…”

“…Baculoviruses have been shown to transduce mammalian cells in vivo, eg in carotid artery, liver, skeletal muscle and murine brain. [1][2][3][4] However, the tropism of baculoviruses in brain has remained unclear. Knowledge about the cell-type-dependent susceptibility for transduction can be utilized for targeting of transgene into certain cell types.…”

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

“…They have been shown to transduce various cell types in vitro and in vivo with significant efficiency. [2][3][4][5] However, efficient gene delivery still remains a challenge for baculovirus as for other gene therapy vectors in many target cells. Efficient transduction would allow high transgene expression with a lower viral load.…”

SPECT/CT imaging of baculovirus biodistribution in rat

“…Intramuscular injections have previously been successfully used in transduction of mouse skeletal muscle with VSV-G displaying baculovirus, 37 and in vaccination studies, 29,38 providing data that mouse muscle might be an attractive target. Yet, in the present study, rat muscle did not result in detectable transduction, as opposed to rabbit muscle, which was transduced with VSV-GED displaying baculovirus.…”