BACKGROUND: Baculovirus, which is widely utilized as an excellent tool for the production of recombinant protein in insect cells, has recently emerged as a novel and attractive gene delivery vehicle for mammalian cells. Alphavirus, such as Semliki Forest virus (SFV), has also received considerable attention for use as expression vectors because of its self-replicating property. In the present study, we investigated the characterization of recombinant baculovirus incorporating a hybrid cytomegalovirus (CMV) promoter/SFV replicon. METHODS: Recombinant baculovirus containing the hybrid CMV promoter/SFV replicon was constructed. Using enhanced green fluorescence protein (EGFP) as the reporter gene, gene delivery efficiencies and the ability to express heterogenous protein in mammalian cells were evaluated. Optimal transduction conditions, including transduction temperature, time and dose, were also investigated. RESULTS: The obtained recombinant baculovirus, Bac-CMV/SFV-EGFP, exhibited high transduction efficiency and high-level expression of reporter protein in mammalian cells. Furthermore, this recombinant baculovirus could induce apoptosis in mammalian cells in the course of transduction, as demonstrated by the observed DNA laddering patterns and increased caspase-3 activity. CONCLUSIONS: The developed baculovirus vector has a high transduction efficiency and the ability to mediate foreign gene expression in mammalian cells. Taken together with its pro-apoptotic properties, this baculovirus vector may provide an alternative tool for vaccine development.
BACKGROUND: Baculovirus, which is widely utilized as an excellent tool for the production of recombinant protein in insect cells, has recently emerged as a novel and attractive gene delivery vehicle for mammalian cells. Alphavirus, such as Semliki Forest virus (SFV), has also received considerable attention for use as expression vectors because of its self-replicating property. In the present study, we investigated the characterization of recombinant baculovirus incorporating a hybrid cytomegalovirus (CMV) promoter/SFV replicon. METHODS: Recombinant baculovirus containing the hybrid CMV promoter/SFV replicon was constructed. Using enhanced green fluorescence protein (EGFP) as the reporter gene, gene delivery efficiencies and the ability to express heterogenous protein in mammalian cells were evaluated. Optimal transduction conditions, including transduction temperature, time and dose, were also investigated. RESULTS: The obtained recombinant baculovirus, Bac-CMV/SFV-EGFP, exhibited high transduction efficiency and high-level expression of reporter protein in mammalian cells. Furthermore, this recombinant baculovirus could induce apoptosis in mammalian cells in the course of transduction, as demonstrated by the observed DNA laddering patterns and increased caspase-3 activity. CONCLUSIONS: The developed baculovirus vector has a high transduction efficiency and the ability to mediate foreign gene expression in mammalian cells. Taken together with its pro-apoptotic properties, this baculovirus vector may provide an alternative tool for vaccine development.
Authors: Kari J Airenne; Yu-Chen Hu; Thomas A Kost; Richard H Smith; Robert M Kotin; Chikako Ono; Yoshiharu Matsuura; Shu Wang; Seppo Ylä-Herttuala Journal: Mol Ther Date: 2013-02-26 Impact factor: 11.454
Authors: April Goehring; Chia-Hsueh Lee; Kevin H Wang; Jennifer Carlisle Michel; Derek P Claxton; Isabelle Baconguis; Thorsten Althoff; Suzanne Fischer; K Christopher Garcia; Eric Gouaux Journal: Nat Protoc Date: 2014-10-09 Impact factor: 13.491
Authors: Sanne Hindriksen; Arne J Bramer; My Anh Truong; Martijn J M Vromans; Jasmin B Post; Ingrid Verlaan-Klink; Hugo J Snippert; Susanne M A Lens; Michael A Hadders Journal: PLoS One Date: 2017-06-22 Impact factor: 3.240