Sustained ex vivo and in vivo transfer of a reporter gene using polyoma virus pseudocapsids

Non-viral gene delivery in skeletal muscle: a protein factory

“…Indeed, whole inactive viral particles or viral proteins, particularly those derived from adenoviruses, have been shown to enhance transgene expression. 72,76,77 Although the results are encouraging, the amount of viral protein required presents an impediment to therapeutic usage of this approach for in vivo transfection. However, it may have some applicability for ex vivo gene therapy.…”

Scanning surface confocal microscopy for simultaneous topographical and fluorescence imaging: Application to single virus-like particle entry into a cell

“…Capsid-like particles were purified by sucrose and CsCl gradient centrifugation from Hi5 insect cells infected with recombinant baculovirus, as described (12), and resuspended in sterile 20 mM Hepes, pH 7.5 at 3-6 mg͞ml. Capsid-like particles were covalently labeled with Cy3 by using the FluoroLink Cy3 monofunctional dye (Amersham Pharmacia).…”

“…Capsid-like particles were covalently labeled with Cy3 by using the FluoroLink Cy3 monofunctional dye (Amersham Pharmacia). Cy3-labeled capsid-like particles were incubated with Qiagen-purified, supercoiled plasmid DNA at a molar ratio of 5:1 to produce VLPs, as described (12). COS 7 cells (5 ϫ 10 4 cells per well) were plated on 13-mm glass coverslips in 24-well plates, incubated overnight at 37°C in DMEM containing 5% (vol͞vol) FCS and then incubated for 30 min at 0°C with VLPs in 400 l of DMEM containing 20 mM Hepes, pH 7.5, 0.5% BSA (0.25 g of plasmid DNA per well).…”

“…We have chosen to use polyoma virus-like particles (VLPs) as a model system to validate our method and to gain insight into their mechanism of entry into cells. Polyoma VLPs are nanospheres composed of 360 molecules of a single viral coat protein, VP1, that have been shown to interact with and carry plasmid DNA into cells (12,13). The protein nanospheres encapsidate 1-2 kb of the plasmid DNA whereas the remainder loosely associates with the outside of the VLP structure (14,15).…”

Efficient gene transfer using the human JC virus-like particle that inhibits human colon adenocarcinoma growth in a nude mouse model

“…Since then, progress has been made in the use of polyomavirus capsid as a gene delivery vector. [18][19][20][21][22][23][24][25][26][27] However, the efficiency of gene transduction using polyomavirus vector is low. In part, this may have been caused by inefficient DNA packaging.…”