Upon entrance into mammalian cells, baculovirus produces little to no microscopically observable cytotoxicity even at a high multiplicity of infection (MOI) ( Kost and Condreay 2002). Unlike many human viruses, baculovirus is incapable of replicating in mammalian cells because of transcriptional silence of its major regulatory genes in these cells. One of our recent studies has demonstrated the effectiveness of using baculoviral vectors for malignant glioma therapy in an animal model ( Wang et al.
As the potential target cells for gene therapy, neurons, the major functional cells in the nervous system, and astrocytes, the most abundant cell type in the brain, have drawn continuous research attention.
2006), neurons and astrocytes in the rat brain and spinal cord ( Li et al. 2002), neuronal cells derived from PC12 and C17.2 cell lines ( Li et al. 2000), cuboidal epithelial cells in the adult rat and mouse brain ( Lehtolainen et al. Among nervous system cells, recombinant baculoviral vectors transduce neuroepithelial cells, neuroblastic cells, astroglial cells and neurons in the primary cell cultures of human and rat embryonic brains ( Sarkis et al. The insect baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV)-based vectors have been introduced as an effective delivery vehicle for transgene expression in mammalian cells, both in proliferating and non-proliferating, quiescent cells ( Kost et al. database for annotation, visualization, and integrated discovery.Autographa californica multiple nuclear polyhedrosis virus.These findings should be useful in understanding the molecular basis for neural cell response to baculoviral transduction and in guiding rational therapeutic applications of baculoviral vectors in the central nervous systems. The related genes were mainly those associated with innate immunity, including several of the genes involved in Toll-like receptor signaling pathway and cytokine-cytokine receptor interaction. We demonstrated that the transduction induced host antiviral responses as a major reaction in all three types of samples profiled. To characterize host responses to virus challenge, thus verifying the suitability of using baculovirus for the development of gene therapy strategies in the central nervous system, we used cDNA microarray technology to examine in vitro and in vivo global cellular gene expression profiles in the rat brain, cultured human astrocytes and human neuronal cells after viral transduction. The molecular data reported will be useful for comparative analysis of gene regulation in the GH/PRL gene family in teleosts.Recombinant baculoviral vectors efficiently transduce several types of cells in the brain. From a comparative point of view, this study of PRL gene in Sparus auratus, correlates well with those previously published on tilapia and rainbow trout. Transient expression experiments with 5 ′-delection mutants reveals at least three regulatory regions on the sbPRL gene, two with a stimulatory effect on transcription and one with apparent inhibitory effect. CHO culture cells co-transfected with a sbPRL promoter sequence and a sea bream Pit-1 cDNA expression plasmid showed expression of a linked luciferase reporter gene. Analysis of 1.0 kb of the proximal promoter sequence reveals a consensus TATAA box, up to seven (A/T) 3NCAT consensus motifs for binding of the pituitary-specific factor Pit-1 and putative CREB and GATA binding sites. The gene analyzed comprises 3.5 kb of DNA containing five exons as described previously for other fish PRL genes. A sea bream prolactin (sbPRL) gene was isolated using a prolactin cDNA fragment, generated by PCR as a probe.
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