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- W2341822389 abstract "Gene therapy is a fast growing research field offering more advanced and exquisite solutions contributing traditional medicine by not only alleviating the symptoms of disorders but also permanently removing their causes via gene transfer. Baculoviruses are of great interest as gene therapy vectors for they are infective only to arthropods, mainly insects, but can effectively transduce various human cells. Nevertheless, more research is needed to explore the nuclear entry process, biodistribution and properties of baculoviruses in human cells to assure the virus safe for transgene delivery. The aim of this study was to establish the active intake of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), the most studied member of Baculoviridae family, into the nucleus of human hepatoma cells (HepG2), to observe the possible transcription of viral genes ie-1 and ie-2 in human cells, in addition to exposing the subnuclear localisation of baculovirus with respect to several prominent transcription sites. Confocal microscopy studies showed baculovirus capsid accumulation into the nucleus at 4 24 h post transduction (p.t). The RT-PCR analysis of transduced HepG2 cells showed transcription of both immediate early viral genes ie-1 and ie-2 at 6 48 h p.t. When exploring possible transcription sites in the nucleus, baculovirus was found to be associated very close to promyelocytic leukaemia nuclear bodies at 8 24 h p.t. No colocalization was found between the virus capsid and nuclear speckles. Qualitative confocal microscopy experiments with H2B-EYFP histone expression plasmid and cell-permeable DNA probe DRAQ5TM revealed a change in host cell chromatin structure at 24 48 h p.t. Thus, this study provides more insight into baculovirus interactions with HepG2 cells: the nuclear entry, accumulation, localization and virus-induced changes in the host nuclear morphology and transcription of viral genes. Finally, these results clearly indicate that further modification of the vector is needed before more clinical trials." @default.
- W2341822389 created "2016-06-24" @default.
- W2341822389 creator A5015657873 @default.
- W2341822389 date "2007-01-01" @default.
- W2341822389 modified "2023-09-27" @default.
- W2341822389 title "New insight into the interplay of baculovirus and subnuclear structures in HepG2 cells" @default.
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