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Title of Journal: Arch Virol

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Abbravation: Archives of Virology

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Springer Vienna

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DOI

10.1016/0076-6879(92)15078-Q

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1432-8798

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Engineered retroviral viruslike particles for rec

Authors: Irena Voráčková Pavel Ulbrich William E Diehl Tomáš Ruml
Publish Date: 2013/10/17
Volume: 159, Issue: 4, Pages: 677-688
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Abstract

Retroviral gag proteins as well as fragments minimally containing the capsid CA and nucleocapsid NC subunits of Gag are able to spontaneously assemble into viruslike particles VLPs This occurs in mammalian and bacterial cells as well as in in vitro systems In every circumstance nucleic acids are incorporated into the forming particles Here we took advantage of an in vitro system for the generation of nonenveloped MasonPfizer monkey virus MPMV VLPs derived from a selfassembling CANC subunit of Gag These VLPs were modified through Nterminal extension of CANC with short oligopeptides that after the assembly process were exposed on the surface of VLPs The employed Nterminal modifications allowed specific interaction with target cells expressing prostatespecific membrane antigen Using this system we were able to incorporate selected siRNA into forming VLPs and deliver it into the cytosol of target cells In comparison with other viral vectors designed for targeted transgene delivery this MPMV VLP system represents the lowest risk of generating virusassociated pathology as the VLPs do not contain any viral coding sequences and are formed in a cellfree systemWe would like to thank Michaela Rumlova for kindly providing expression plasmids and polyclonal anticapsid protein antibody and Jan Konvalinka for kindly providing stably transfected HEK 293T cells This research was supported financially by Czech Science Foundation grant P302/12/1895 and Czech Ministry of Education grant LH12011

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