Document Detail


Retroviral vector performance in defined chromosomal Loci of modular packaging cell lines.
MedLine Citation:
PMID:  20222806     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The improvement of safety and titer of retroviral vectors produced in standard retroviral packaging cell lines is hampered because production relies on uncontrollable vector integration events. The influences of chromosomal surroundings make it difficult to dissect the performance of a specific vector from the chromosomal surroundings of the respective integration site. Taking advantage of a technology that relies on the use of packaging cell lines with predefined integration sites, we have systematically evaluated the performance of several retroviral vectors. In two previously established modular packaging cell lines (Flp293A and 293 FLEX) with single, defined chromosomal integration sites, retroviral vectors were integrated by means of Flp-mediated site-specific recombination. Vectors that are distinguished by different long terminal repeat promoters were introduced in either the sense or reverse orientation. The results show that the promoter, viral vector orientation, and integration site are the main determinants of the titer. Furthermore, we exploited the viral production systems to evaluate read-through activity. Read-through is thought to be caused by inefficient termination of vector transcription and is inherent to the nature of retroviral vectors. We assessed the frequency of transduction of sequences flanking the retroviral vectors from both integration sites. The approach presented here provides a platform for systematic design and evaluation of the efficiency and safety of retroviral vectors optimized for a given producer cell line.
Authors:
L Gama-Norton; S Herrmann; R Schucht; A S Coroadinha; R Löw; P M Alves; C C Bartholomae; M Schmidt; C Baum; A Schambach; H Hauser; D Wirth
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Human gene therapy     Volume:  21     ISSN:  1557-7422     ISO Abbreviation:  Hum. Gene Ther.     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-08-11     Completed Date:  2011-01-18     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9008950     Medline TA:  Hum Gene Ther     Country:  United States    
Other Details:
Languages:  eng     Pagination:  979-91     Citation Subset:  IM    
Affiliation:
Helmholtz Center for Infection Research (HZI), 38124 Braunschweig, Germany.
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MeSH Terms
Descriptor/Qualifier:
Cell Line / metabolism*
Chromosomes*
Gene Targeting
Gene Therapy / methods
Genetic Loci*
Genetic Vectors / metabolism*
Promoter Regions, Genetic
Retroviridae / genetics*,  physiology
Transduction, Genetic
Virus Assembly*
Virus Integration

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


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