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Long-term transgene expression in mouse neural progenitor cells modified with phiC31 integrase.
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Long-term transgene expression in mouse neural progenitor cells modified with phiC31 integrase.

Author: A Keravala Affiliation: Department of Genetics, Stanford University School of Medicine, M-334, 300 Pasteur Drive, Stanford, CA 94305-5120, United States.; BK Ormerod; TD Palmer; MP Calos
Edition/Format: Article Article : English
Publication:Journal of neuroscience methods, 2008 Aug 30; 173(2): 299-305
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Summary:
Stem cells can potentially be utilized in combined gene/cell therapies for neural diseases. We examined the ability of the non-viral phiC31 integrase system to promote stable transgene expression in mouse neural progenitor cells (mNPCs). phiC31 integrase catalyzes the sequence-specific integration of attB-containing plasmids into pseudo attP sites in mammalian genomes, to produce long-term transgene expression. We  Read more...
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Document Type: Article
All Authors / Contributors: A Keravala Affiliation: Department of Genetics, Stanford University School of Medicine, M-334, 300 Pasteur Drive, Stanford, CA 94305-5120, United States.; BK Ormerod; TD Palmer; MP Calos
ISSN:0165-0270
Language Note: English
Unique Identifier: 264670766
Awards:

Abstract:

Stem cells can potentially be utilized in combined gene/cell therapies for neural diseases. We examined the ability of the non-viral phiC31 integrase system to promote stable transgene expression in mouse neural progenitor cells (mNPCs). phiC31 integrase catalyzes the sequence-specific integration of attB-containing plasmids into pseudo attP sites in mammalian genomes, to produce long-term transgene expression. We achieved gene transfer by co-nucleofection of a plasmid carrying the luciferase marker gene and an attB site and a plasmid expressing integrase in mNPCs that had been generated in a neurosphere preparation. Luciferase expression was quantified in live cells for 8 weeks, revealing persistence of gene expression. Sequence-specific integration at a preferred pseudo attP site in the mouse genome was detected by using PCR. Furthermore, sustained transgene expression was demonstrated in genetically modified NPCs that were cultured in conditions that promoted either growth or differentiation into neurons and astrocytes. Our results demonstrate that the phiC31 integrase system produces stable transgene expression in adult mNPCs and their progeny and may be useful in strategies for combating neurodegenerative disorders.

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