Document Detail


Cre/loxP-mediated adenovirus type 5 packaging signal excision demonstrates that core element VI is sufficient for virus packaging.
MedLine Citation:
PMID:  12758179     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Previous analyses have demonstrated that packaging of the adenovirus type 5 (Ad5) genome is dependent on at least seven cis-acting elements, called AI to AVII, which are located in the left-end region of the genome. These elements have different packaging efficiencies, and without AI through AV, viral DNA cannot be packaged. Here we report the identification of the cis-acting Ad5 packaging domain in vivo by using the Cre/loxP system. We found that an adenoviral DNA fragment (nt 192 to nt 358), which includes elements AI to AV, is excised by Cre recombinase and packaged into capsids. Furthermore, this mutant adenovirus replicated so efficiently by repetitive propagation that its purification by CsCI equilibrium gradient was possible. This study clarified that the region from nt 358 to nt 454 on the viral genome is sufficient for packaging. Recently, the helper-dependent adenoviral vector (HDAd) construction system has been developed for the purpose of gene therapy. This system uses a helper virus with two parallel loxP sites flanking the packaging signal. This region is eliminated by Cre-mediated excision, which prevents helper virus packaging. Our data provide useful information regarding factors affecting efficient elimination.
Authors:
Yasushi Maeda; En Kimura; Yuji Uchida; Yasuto Nishida; Satoshi Yamashita; Toshiyuki Arima; Makoto Uchino
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Virology     Volume:  309     ISSN:  0042-6822     ISO Abbreviation:  Virology     Publication Date:  2003 May 
Date Detail:
Created Date:  2003-05-21     Completed Date:  2003-07-07     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0110674     Medline TA:  Virology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  330-8     Citation Subset:  IM    
Affiliation:
Department of Neurology, Kumamoto University, School of Medicine, 1-1-1 Honjo, Kumamoto, Kumamoto 860-0811, Japan. yasushi-maeda@fc.kuh.kumamoto-u.ac.jp
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MeSH Terms
Descriptor/Qualifier:
Adenoviruses, Human / chemistry,  genetics,  metabolism*,  physiology
Animals
Base Sequence
COS Cells
Cell Line
DNA, Viral / metabolism
Enhancer Elements, Genetic* / genetics*
Genome, Viral
Humans
Integrases / metabolism*
Molecular Sequence Data
Sequence Deletion
Signal Transduction*
Viral Proteins / metabolism*
Virus Assembly*
Chemical
Reg. No./Substance:
0/DNA, Viral; 0/Viral Proteins; EC 2.7.7.-/Cre recombinase; EC 2.7.7.-/Integrases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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