Cellular contaminants of adeno-associated virus vector stocks can enhance transduction

Tenenbaum, Liliane; Hamdane, Malika; Pouzet, Marianne; Avalosse, Bernard; Stathopoulos, Angelike; Jurysta, Fabrice; Rosenbaum, Claudia; Hanemann, C. Oliver; Levivier, Marc; Velu, Thierry

doi:10.1038/sj.gt.3300904

Cited by 24 publications

(23 citation statements)

References 41 publications

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“…The mechanism for this difference is currently unclear, however, it is possible that it may be related to batch-to-batch variability in contaminants of the viral preparation. 22 Nevertheless, the major point in our experiments is that RA/TSA enhance and delay transgene expression versus control regardless of the precise timing of peak ␤-gal activity. Taken together, the results suggest that the progressive reduction of transgene expression that follows AdV-mediated gene transfer into mammalian cells both in vitro and in vivo, is dependent on transcriptional silencing processes sensitive to HAT and HDAC activity.…”

Section: Ra and Tsa Effects On Transgene Expression In Vivomentioning

confidence: 99%

Transcriptionally active drugs improve adenovirus vector performance in vitro and in vivo

et al. 2000

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Section: Ra and Tsa Effects On Transgene Expression In Vivomentioning

confidence: 99%

Transcriptionally active drugs improve adenovirus vector performance in vitro and in vivo

et al. 2000

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“…on April 1, 2019 by guest http://jvi.asm.org/ rAAV may nonspecifically trap transgene products, as previously suggested (4,65). As shown in Fig.…”

Section: Vol 74 2000 Aav Transduction Of Human Skeletal Muscle Cellmentioning

confidence: 57%

Kinetics of Recombinant Adeno-Associated Virus-Mediated Gene Transfer

Malik

Monahan

Allen

et al. 2000

J Virol

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Recombinant adeno-associated virus (rAAV) vectors have been shown to be useful for efficient gene delivery to a variety of dividing and nondividing cells. Mechanisms responsible for the long-term, persistent expression of the rAAV transgene are not well understood. In this study we investigated the kinetics of rAAV-mediated human factor IX (hFIX) gene transfer into human primary myoblasts and myotubes. Transduction of both myoblasts and myotubes occured with a similar and high efficiency. After 3 to 4 weeks of transduction, rAAV with a cytomegalovirus (CMV) promoter showed 10-to 15-fold higher expression than that with a musclespecific creatine kinase enhancer linked to ␤-actin promoter. Factor IX expression from transduced myoblasts as well as myotubes reached levels as high as approximately 2 g of hFIX/10 6 cells/day. Southern blot analyses of high-molecular-weight (HMW) cellular genomic and Hirt DNAs isolated from rAAV/CMVhFIXm1-transduced cells showed that the conversion of single-stranded vector genomes to double-stranded DNA forms, but not the level of the integrated forms in HMW DNA, correlated with increasing expression of the transgene. Together, these results indicate that rAAV can transduce both proliferating and terminally differentiated muscle cells at about the same efficiency, that expression of transgenes increases linearly over their lifetime with no initial lag phase, and that increasing expression correlates with the appearance of double-stranded episomal rAAV genomes. Evidence showing that the rAAV virions can copackage hFIX, presumably nonspecifically, was also obtained.

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“…The pTR-enhanced GFP (EGFP) plasmid that contains the EGFP gene (Clonetech, Montigny-leBretonneux, France) under the control of cytomegalovirus (CMV) promoter and the pTet bidi vector containing the EGFP reporter gene and the reverse tetracycline transactivator (rtTA) under the control of a central and bidirectional tetracycline-responsive promoter containing seven tetracycline operators have been described previously [7,8]. The pTR-MBP-EGFP plasmid was derived from pCMV-EGFP [8] by replacing the CMV promoter by a 0.87 kb fragment of the myelin basic protein (MBP) promoter which was polymerase chain reaction-amplified from genomic mouse DNA, using SpeI and ApaI sites. The pTR-UF12.1 plasmid containing GFP (F64L,S65T) under the control of the hybrid CMV immediate early enhancer/chicken b-actin (CBA) promoter/exon 1/intron [9] was a kind gift from Dr S. Zolotukhin (University of Florida).…”

Section: Vectorsmentioning

confidence: 99%

“…To assay the potential of AAV-2 for ex-vivo gene transfer in the CNS, SCs were transplanted in lysophosphatidylcholineinduced demyelinated lesions known to elicit their differentiation in myelin-forming cells in the mouse spinal cord [6][7][8][9][10][11][12][13]. Compared with uninfected SCs, the number of HN + surviving SCs was reduced after infected cell transplantation (Po0.01).…”

Section: Potential Of Adeno-associated Virus Type-2 For Transgene Expmentioning

confidence: 99%

Efficiency of adeno-associated virus type-2 vectors in non-human primate Schwann cells

et al. 2005

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Adult macaque Schwann cells were infected using adeno-associated virus type-2-derived vectors expressing the green fluorescent protein reporter gene under the control of the cytomegalovirus, the hybrid cytomegalovirus-betaactin, the myelin basic protein or the tetracycline-inducible promoters. On the basis of green fluorescent protein expression, gene transfer efficiency was compared in resting and dividing conditions following or not following hydroxyurea or etoposide treatment. Hydroxyurea allowed promoter-dependent expression of green fluorescent protein in infected Schwann cells. Etoposide treatment led to a high percentage of green fluorescent protein expressing cells (over 50%) with all promoters tested. When infected cells were grafted into demyelinated nude mice spinal cord, green fluorescent protein expression was only observed with the cytomegalovirus-betaactin and tetracycline-inducible promoters. In addition, adeno-associated virus type-2 infection reduced the grafted cell survival but increased their differentiation.

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Cellular contaminants of adeno-associated virus vector stocks can enhance transduction

Cited by 24 publications

References 41 publications

Transcriptionally active drugs improve adenovirus vector performance in vitro and in vivo

Transcriptionally active drugs improve adenovirus vector performance in vitro and in vivo

Kinetics of Recombinant Adeno-Associated Virus-Mediated Gene Transfer

Efficiency of adeno-associated virus type-2 vectors in non-human primate Schwann cells

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