Efficiency of delivery of DNA to cells by bovine papillomavirus type-1 L1/L2 pseudovirions

• Published in: Applied microbiology and biotechnology Vol. 56; no. 1/2; pp. 150 - 156
• Main Authors: Liu, Y, Frazer, I.H, Liu, W.J, Liu, X.S, McMillan, N, Zhao, K.N
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.07.2001; Springer Nature B.V
• Subjects: Animals; antiserum; Biological and medical sciences; Biotechnology; Bovine papillomavirus; Bovine papillomavirus 1; Bovine papillomavirus 1 - physiology; Capsid - physiology; Capsid Proteins; Cattle; COS Cells; Deoxyribonuclease; deoxyribonucleases; Deoxyribonucleic acid; DNA; DNA - metabolism; Efficiency; Encapsidation; Fluorescence; Fundamental and applied biological sciences. Psychology; Gene therapy; Gene transfer; Green fluorescent protein; Health. Pharmaceutical industry; Industrial applications and implications. Economical aspects; Infectivity; Packaging; Papilloma viruses; Papillomavirus; pathogenicity; Photovoltaic cells; Plasmids; production technology; Proteins; Reporter gene; reporter genes; Vaccinia virus; Virion - physiology; Virus Assembly; Virus-like particles; Viruses; Cell culture; Pseudovirion; Monkey; Papovaviridae; Kidney; Genetic transfer; Papillomavirus; Virus; Plasmid; Bovine papillomavirus; Vertebrata; Encapsidation; Cell line; Mammalia; Animal; DNA; Primates; Bovine papillomavirus 1; Gene therapy; Vector
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s002530100655

To investigate the efficiency of encapsidation of plasmid by papillomavirus virus-like particles (PV VLPs), and the infectivity of the resultant PV pseudovirions, Cos-1 cells were transfected with an 8-kb plasmid incorporating a green fluorescent protein (GFP) reporter gene (pGSV), and infected with bovine PV (BPV-1) L1/L2 recombinant vaccinia virus to produce BPV1 pseudovirions. Approximately 1 in 1.5x10(4) of dense (1.35 g/ml) PV pseudovirions and 0.3 in 10(4) of less-dense (1.29 g/ml) pseudovirions packaged an intact pGSV plasmid. The majority (>75%) of packaged plasmids contained deletions, and the deletions affected all tested genes. After exposure of Cos-1 cells to BPV-1 pseudovirions at an MOI of 40,000:1, 6% of cells expressed GFP, giving a calculated efficiency of delivery of the pGSV plasmid, by pseudovirions which had packaged an intact plasmid, of approximately 5%. Plasmid delivery was not effected by purified pGSV plasmid, was blocked by antiserum against BPV-1, and was not blocked by DNase treatment of pseudovirions, confirming that delivery was mediated by DNA within the pseudovirion. We conclude that a major limitation to the use of PV pseudovirions as a gene delivery system is that intact plasmid DNA is not efficiently selected for packaging by VLPs in cell-based pseudovirions production systems.