Retroviral vector performance in defined chromosomal Loci of modular packaging cell lines.
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Coroadinha, A S
Alves, P M
Bartholomae, C C
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AbstractThe 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.
CitationRetroviral vector performance in defined chromosomal Loci of modular packaging cell lines. 2010, 21 (8):979-91 Hum. Gene Ther.
AffiliationHelmholtz Center for Infection Research (HZI), 38124 Braunschweig, Germany.
JournalHuman gene therapy
The following license files are associated with this item:
- A new generation of retroviral producer cells: predictable and stable virus production by Flp-mediated site-specific integration of retroviral vectors.
- Authors: Schucht R, Coroadinha AS, Zanta-Boussif MA, Verhoeyen E, Carrondo MJ, Hauser H, Wirth D
- Issue date: 2006 Aug
- Evaluation of retroviral vector design in defined chromosomal loci by Flp-mediated cassette replacement.
- Authors: Verhoeyen E, Hauser H, Wirth D
- Issue date: 2001 May 20
- The use of recombinase mediated cassette exchange in retroviral vector producer cell lines: predictability and efficiency by transgene exchange.
- Authors: Coroadinha AS, Schucht R, Gama-Norton L, Wirth D, Hauser H, Carrondo MJ
- Issue date: 2006 Jul 13
- Integrated strategy for the production of therapeutic retroviral vectors.
- Authors: Carrondo M, Panet A, Wirth D, Coroadinha AS, Cruz P, Falk H, Schucht R, Dupont F, Geny-Fiamma C, Merten OW, Hauser H
- Issue date: 2011 Mar
- Retroviral vectors produced in the cytoplasmic vaccinia virus system transduce intron-containing genes.
- Authors: Konetschny C, Holzer GW, Falkner FG
- Issue date: 2002 Feb