| pubmed-article:14671083 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C0007634 | lld:lifeskim |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C0282641 | lld:lifeskim |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C0017262 | lld:lifeskim |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C1314677 | lld:lifeskim |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C0443252 | lld:lifeskim |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C0127400 | lld:lifeskim |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C1514485 | lld:lifeskim |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C2911684 | lld:lifeskim |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C1510800 | lld:lifeskim |
| pubmed-article:14671083 | lifeskim:mentions | umls-concept:C0185117 | lld:lifeskim |
| pubmed-article:14671083 | pubmed:issue | 1 | lld:pubmed |
| pubmed-article:14671083 | pubmed:dateCreated | 2003-12-12 | lld:pubmed |
| pubmed-article:14671083 | pubmed:abstractText | High-capacity "gutless" adenovirus vectors (HC-AdV) mediate long-term transgene expression in resting cells in vitro and in vivo because of low toxicity and immunogenicity. However, in proliferating cells, expression is transient since HC-AdV genomes do not possess elements that allow for replication and segregation of the replicated genomes to daughter cells. We developed a binary HC-AdV system that, under certain conditions, allows for significantly prolonged episomal maintenance of HC-AdV genomes in proliferating tissue culture cells, resulting in sustained transgene expression. After transduction of target cells the linear HC-AdV genomes were circularized by the DNA recombinase FLPe, which was expressed from the second HC-AdV. The oriP/EBNA-1 replication system derived from Epstein-Barr virus, as well as the human replication origin from the lamin B2 locus, were used as cis elements to test for replication of the 28-kb circular vector genomes with or without selective pressure. Depending on the system, up to 98% of the circularized genomes were replicated and segregated to daughter cells, as demonstrated by Southern assays and as confirmed by monitoring EGFP transgene expression. Surprisingly, in the absence of FLPe recombinase, a small but significant number of HC-AdV genomes spontaneously circularized after transduction of target cells. These circles, found to contain end-to-end joined adenovirus termini, replicated with increased efficiency compared to vectors circularized by FLPe. After further improvements, this HC-AdV system might be suitable for gene therapy applications requiring long-term transgene expression. | lld:pubmed |
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| pubmed-article:14671083 | pubmed:language | eng | lld:pubmed |
| pubmed-article:14671083 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:14671083 | pubmed:citationSubset | IM | lld:pubmed |
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| pubmed-article:14671083 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:14671083 | pubmed:month | Jan | lld:pubmed |
| pubmed-article:14671083 | pubmed:issn | 0022-538X | lld:pubmed |
| pubmed-article:14671083 | pubmed:author | pubmed-author:KochanekStefa... | lld:pubmed |
| pubmed-article:14671083 | pubmed:author | pubmed-author:KreppelFloria... | lld:pubmed |
| pubmed-article:14671083 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:14671083 | pubmed:volume | 78 | lld:pubmed |
| pubmed-article:14671083 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:14671083 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:14671083 | pubmed:pagination | 9-22 | lld:pubmed |
| pubmed-article:14671083 | pubmed:dateRevised | 2009-11-18 | lld:pubmed |
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| pubmed-article:14671083 | pubmed:year | 2004 | lld:pubmed |
| pubmed-article:14671083 | pubmed:articleTitle | Long-term transgene expression in proliferating cells mediated by episomally maintained high-capacity adenovirus vectors. | lld:pubmed |
| pubmed-article:14671083 | pubmed:affiliation | Center for Molecular Medicine Cologne, University of Cologne, D-50931 Cologne, Germany. | lld:pubmed |
| pubmed-article:14671083 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:14671083 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
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