This chapter describes the protocol for the preparation of recombinant adenoviruses and infection of target cells to transiently express G protein-coupled receptors (GPCRs) or other proteins of interest. Adenoviruses are non-enveloped viruses containing a linear double-stranded DNA genome. Their life cycle does not normally involve integration into the host genome, rather they replicate as episomal -elements in the nucleus of the host cell, and consequently there is no risk of insertional mutagenesis. Up to 30 kb out of the 35 kb of the wild-type adenovirus genome can be replaced by foreign DNA. Adenoviral vectors are very efficient in transducing target cells in vitro and in vivo and can be produced at high titers (>10 11/mL). The viral infection has a number of useful features: (1) the efficiency of gene transduction is very high (up to 100% in sensitive cells); (2) the infection is easy and does not physically alter the cell membrane for gene transduction; (3) it is possible to infect cells that are resistant to transfection with plasmids (including nondividing cells); and (4) the viral vectors can be used for infection in vivo (including gene therapy) and can potentially be targeted cell-specifically.
Viral Infection for GPCR Expression in Eukaryotic Cells / Antonio, Porcellini; Iacovelli, Luisa; DE BLASI, Antonio. - 746(2011), pp. 39-51. - METHODS IN MOLECULAR BIOLOGY. [10.1007/978-1-61779-126-0_3].
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Titolo: | Viral Infection for GPCR Expression in Eukaryotic Cells | |
Autori: | ||
Data di pubblicazione: | 2011 | |
Serie: | ||
Citazione: | Viral Infection for GPCR Expression in Eukaryotic Cells / Antonio, Porcellini; Iacovelli, Luisa; DE BLASI, Antonio. - 746(2011), pp. 39-51. - METHODS IN MOLECULAR BIOLOGY. [10.1007/978-1-61779-126-0_3]. | |
Handle: | http://hdl.handle.net/11573/443009 | |
ISBN: | 9781617791253 9781617791260 | |
Appartiene alla tipologia: | 02a Capitolo o Articolo |