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Viral Infection for GPCR Expression in Eukaryotic Cells.
MedLine Citation:
PMID:  21607851     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
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.
Authors:
Antonio Porcellini; Luisa Iacovelli; Antonio De Blasi
Publication Detail:
Type:  JOURNAL ARTICLE    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  746     ISSN:  1940-6029     ISO Abbreviation:  -     Publication Date:  2011  
Date Detail:
Created Date:  2011-5-24     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  39-51     Citation Subset:  -    
Affiliation:
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