Insect-derived cell lines are used extensively to produce recombinant proteins because they are capable of performing a range of post-translational modifications. Due to their significance in biotechnological applications, various methods have been developed to transfect them. In this study, we introduce a virosome constructed from vesicular stomatitis virus (VSV) as a new delivery system for sf9 cells. We labeled these VSV virosomes by fluorescent probe Rhodamine B chloride (R18). By fluorescence microscope observation and conducting a fusion assay, we confirmed the uptake of VSV virosomes via endocytosis by sf9 cells and their fusion with the endosomal membrane. Moreover, we incubated cationic VSV virosomes with a GFP-expressing bacmid and transfected sf9 cells, after 24 h some cells expressed GFP indicating the ability of VSV virosomes to deliver heterologous DNA to these cells. This is the first report of a virosome-based delivery system introduced for an insect cell line.
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http://dx.doi.org/10.3109/08982104.2016.1144205 | DOI Listing |
Curr Issues Mol Biol
January 2025
Department of Biopharmacy, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China.
(Mhp) infection severely affects the daily weight gain and feed-to-meat ratio of pigs, while secondary infections with other pathogens can further lead to increased mortality, causing significant economic losses to the pig industry. CD40L is a molecular adjuvant that enhances the cellular and humoral immune responses to vaccines. In this study, the CD40L peptide was fused to the C-terminus of the chimeric P97R1P46P42 protein by genetic engineering using the pFastBac Dual vector.
View Article and Find Full Text PDFCurr Pharm Biotechnol
January 2025
College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310000, China.
Background: Gene therapy has been effectively applied in many biological studies and for the treatment of many genetic or cancer diseases. Currently, Recombinant Adeno- Associated Viruses (rAAVs) are one of the main types of delivery vectors used for gene therapy. rAAV vectors produced via the Sf9 cells have the advantages of high rAAV yields, easy scaleup, and low cost.
View Article and Find Full Text PDFBiotechnol Bioeng
January 2025
Instituto de Biologia Experimental e Tecnológica (iBET), Oeiras, Portugal.
The insect cell-baculovirus expression vector system (IC-BEVS) has been an asset to produce biologics for over 30 years. With the current trend in biotechnology shifting toward process intensification and integration, developing intensified processes such as continuous production is crucial to hold this platform as a suitable alternative to others. However, the implementation of continuous production has been hindered by the lytic nature of this expression system and the process-detrimental virus passage effect.
View Article and Find Full Text PDFJ Immunol Methods
January 2025
ICAR-Indian Veterinary Research Institute, Bangalore, Karnataka 560024, India.
Bluetongue (BT) is a vector-borne viral disease of multiple domestic and wild ruminants across the globe. The VP7 protein of bluetongue virus (BTV) is the major immune-dominant structural protein that is conserved across the BTV serotypes and therefore, targeted for the development of immuno-diagnostics for BT. In this study, full-length recombinant VP7 protein (rVP7) of BTV-1 was expressed in Trochoplusia ni derived insect cells (Tn5) using codon-optimized synthetic gene construct through baculovirus expression system.
View Article and Find Full Text PDFMol Biol Rep
January 2025
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK.
Background: Ribosomal protein S6 kinase 1 (p70S6K1) is a member of the AGC family of serine/threonine kinases which plays a role in various cellular processes, including protein synthesis, cell growth, and survival. Dysregulation of p70S6K1, characterized by its overexpression and/or hyperactivation, has been implicated in numerous human pathologies, particularly in several types of cancer. Therefore, generating active, recombinant p70S6K1 is critical for investigating its role in cancer biology and for developing novel diagnostic or therapeutic approaches.
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