Site-specific integration by adeno-associated virus.

Kotin, Robert M.; Siniscalco, M.; Samulski, R. Jude; Zhu, X. D.; Hunter, L.; Laughlin, C. A.; McLaughlin, S.; Muzyczka, N.; Rocchi, M.; Berns, Kenneth I.

doi:10.1073/pnas.87.6.2211

Cited by 847 publications

(546 citation statements)

References 12 publications

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“…Although the wild-type virus efficiently integrates in a specific location in the human genome (AAVS1 on chromosome 19), 29 integration of rAAV genomes is typically passive, of low frequency and largely random. [30][31][32] The vast majority of rAAV vector genomes remain as episomes in the host cell nucleus, with only an estimated 0.5% of genomes integrating into the host genome, 33 probably at pre-existing chromosomal breaks.…”

Section: Introductionmentioning

confidence: 99%

Absence of ocular malignant transformation after sub-retinal delivery of rAAV2/2 or integrating lentiviral vectors in p53-deficient mice

Balaggan

Durán²,

Georgiadis³

et al. 2011

Gene Ther

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Insertional mutagenesis following gene therapy with gammaretroviral vectors can cause the development of lymphoproliferation in children with X-linked severe combined immunodeficiency. In experimental studies, recombinant adeno-associated virus (rAAV) vectors have also been reported to increase susceptibility to carcinogenesis. The possibility of vector-induced transformation in quiescent ocular cells is probably significantly lower than in mitotically active cells, but given the increasing number of clinical applications of rAAV and lentiviral vectors for ocular disease, a specific assessment of their oncogenic potential in the eye is important. In this study, we investigated the effect of rAAV2/2 and integrating HIV-1 vectors upon the incidence of ocular neoplasia in p53 tumour-suppressor gene-knockout (p53 À/À ) mice, which are highly susceptible to intraocular malignant transformation. Subretinal injections of high titre rAAV2/2 or integrating HIV-1 vectors induced no tumours in p53 À/À or p53 +/À animals, nor significantly affected their natural longevity. We conclude that any insertional events arising from subretinal delivery of these vectors appear insufficient to cause intraocular malignancy, even in highly susceptible animals. These findings support the continued development of these vectors for ocular applications.

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Section: Introductionmentioning

confidence: 99%

Absence of ocular malignant transformation after sub-retinal delivery of rAAV2/2 or integrating lentiviral vectors in p53-deficient mice

Balaggan

Durán²,

Georgiadis³

et al. 2011

Gene Ther

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“…6 -8 However, the packaging capacity of recombinant AAVs is severely restricted owing to the small size of the viral genome; in addition, although integration of wt AAV DNA occurs in a site-specific manner, recombinant AAV vectors integrate randomly. 9,10 An ideal stem cell vector would share the efficient delivery characteristics of the above viruses, but should also be capable of accommodating larger therapeutic genes and be able to maintain itself in a dividing cell population without the need for integration. Herpesviruses are large DNA viruses possessing genomes of between 100 and 250 kb.…”

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confidence: 99%

A herpesvirus saimiri-based gene therapy vector with potential for use in cancer immunotherapy

et al. 2000

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The herpesvirus saimiri (HVS) genome has the capacity to incorporate large amounts of heterologous DNA and can be maintained episomally in many different human cell types. To evaluate the efficacy of HVS-mediated gene transfer into human hemopoietic cells, we investigated the ability of an HVS-based construct, carrying the enhanced green fluorescent protein (EGFP) and neomycin resistance genes, to transduce a variety of human hemopoietic cell lines and primary CD34 ϩ cells. As measured by flow cytometry, the numbers of EGFP ϩ cells at 2 days postinfection differed between various cell types ranging, from 1.3% for KG1 cells to 56.8% for THP-1 cells. In addition, the expression of EGFP in Jurkat cells was retained at Ͼ95% per round of cell division over a period of 6 weeks (comparable with Epstein-Barr virus-derived gene therapy systems). Although the virus was not specifically disabled, no lytic viral mRNAs could be detected in transduced Jurkat cells, and infectious virus could not be detected by sensitive virus recovery assay. We also describe a simple centrifugation method that increases the efficiency of transduction by Ͼ100% in some cases and may be generally applicable to other herpesvirus-based vectors for ex vivo gene delivery. Using this technique, we were able to demonstrate a tropism for CD34 ϩ /CD14 ϩ cells, transducing 30% of the population. These cells are known to give rise to dendritic cells (the most potent of the antigen-presenting cells), suggesting that the vector could be used to deliver DNA sequences encoding tumor antigens for cancer immunotherapy.

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“…44 Recent studies from our group and collaborators demonstrate that by flanking the transgene in amplicon vectors with the ITR elements and a Rep expression cassette from AAV, site-specific integration of the transgene at the AAVS1 chromosomal site can be achieved in about 10-30% of transduced human cells, 45,46 as predicted from studies of AAV. 44,47 We have now generated transgenic mice bearing the human AAVS1 locus, 48 which can be crossed to A-T heterozygous mice to create ATM K-O mice with AAVS1 sites, as a model of the human genome.…”

Section: Discussionmentioning

confidence: 99%

HSV-1 amplicon vector-mediated expression of ATM cDNA and correction of the ataxia–telangiectasia cellular phenotype

et al. 2003

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Site-specific integration by adeno-associated virus.

Abstract: Cellular sequences flanking integrated copies of the adeno-associated virus (AAV) genome were isolated from a latently infected clonal human cell line and used to probe genomic blots derived from an additional 21 independently derived clones of human cells latently infected with AAV.

Cited by 847 publications

References 12 publications

Absence of ocular malignant transformation after sub-retinal delivery of rAAV2/2 or integrating lentiviral vectors in p53-deficient mice

Absence of ocular malignant transformation after sub-retinal delivery of rAAV2/2 or integrating lentiviral vectors in p53-deficient mice

A herpesvirus saimiri-based gene therapy vector with potential for use in cancer immunotherapy

HSV-1 amplicon vector-mediated expression of ATM cDNA and correction of the ataxia–telangiectasia cellular phenotype

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