Sustained ex vivo and in vivo transfer of a reporter gene using polyoma virus pseudocapsids.
Properties of a virus-like artificial gene delivery vehicle, synthesised from recombinant major coat protein of mouse polyoma virus, have been explored. The protein, VP1, self assembles into protein spheres, or 'pseudocapsids, which can bind and transfer DNA into cells in vitro and in vivo. Here, the ability of pseudocapsids to carry DNA into a complex cell system (ex vivo organ cultures of rabbit cornea) or whole animals (mice) has been assessed. Evidence from histochemical and PCR experiments indicate that pseudocapsids stimulate uptake and stable maintenance of marker DNA in nondividing corneal cells as efficiently as a recombinant adenovirus. In athymic and immunocompetent mice, gene transmission occurs with no apparent adverse effects on the animals. In the presence of pseudocapsids, the marker gene was transferred to a range of organs, including the brains of animals, following peripheral or intranasal administration. In immunocompetent mice, significant long-term transcriptional expression (at least 22 weeks) was observed with pseudocapsids, a period significantly longer than observed with DNA alone (several weeks only), again with no obvious adverse effects. This study demonstrates that pseudocapsids from the murine virus, polyoma, constitute a novel transfer agent for long-term gene therapeutic applications in tissues or whole animals.
Department of Infectious Diseases (Virology), Imperial College School of Medicine at Hammersmith Hospital, London, UK.
N
N
Krauzewicz
C
C
Cox
E
E
Soeda
B
B
Clark
S
S
Rayner
B E
BE
Griffin
D000818:Animals; D002213:Capsid; D036022:Capsid Proteins; D003315:Cornea / Q000201:enzymology; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D016133:Polymerase Chain Reaction; D011120:Polyomavirus; D011817:Rabbits; D013997:Time Factors; D001616:beta-Galactosidase / Q000235:genetics
D036022:Capsid Proteins; C062481:VP1 protein, polyomavirus; D001616:beta-Galactosidase