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    Polyplex-mediated gene transfer into human retinal pigment epithelial cells in vitro

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    Authors
    Chaum, Edward
    Hatton, Mark P.
    Stein, Gary
    UMass Chan Affiliations
    Department of Cell Biology
    Department of Medicine, Division of Ophthalmology
    Document Type
    Journal Article
    Publication Date
    1999-12-01
    Keywords
    DNA
    Gene Expression
    *Gene Transfer Techniques
    Humans
    In Vitro Techniques
    Pigment Epithelium of Eye
    Transgenes
    Biochemistry
    Cell Biology
    Eye Diseases
    Ophthalmology
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    Link to Full Text
    http://dx.doi.org/10.1002/(SICI)1097-4644(20000101)76:1%3C153::AID-JCB15%3E3.0.CO;2-%23
    Abstract
    The human retinal pigment epithelium (RPE) is a potential target tissue for directed transfer of candidate genes to treat age-related macular degeneration (AMD). The RPE is uniquely suited to gene therapy protocols that use liposome-mediated DNA transfer because of its high intrinsic phagocytic function in vivo. In these studies, we examined the efficacy of human RPE cell uptake and expression of the green fluorescent protein (GFP) and neomycin resistance marker genes by polyplex-mediated gene transfer in vitro. The effects of varying DNA and polyplex concentration and ratios on GFP transgene expression were examined. A narrow range of experimental conditions were found to maximize transgene expression; most important were the DNA concentration and the DNA:polyplex ratio. The transfection efficiency for human RPE cells was reproducibly 20% in vitro by this method and reached a maximum level of expression after 48 h. There was a rapid decline in gene expression over 2 weeks following polyplex-mediated gene transfer, but stable integration does occur at low frequencies with and without selection.
    Source
    J Cell Biochem. 1999 Nov;76(1):153-60. DOI:10.1002/(SICI)1097-4644(20000101)76:1%3C153::AID-JCB15%3E3.0.CO;2-%23
    DOI
    10.1002/(SICI)1097-4644(20000101)76:1%3C153::AID-JCB15%3E3.0.CO;2-%23
    Permanent Link to this Item
    http://hdl.handle.net/20.500.14038/49228
    PubMed ID
    10581009
    Notes

    Medical student Mark P. Hatton participated in this study as part of the Senior Scholars research program at the University of Massachusetts Medical School.

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    Link to Article in PubMed
    ae974a485f413a2113503eed53cd6c53
    10.1002/(SICI)1097-4644(20000101)76:1%3C153::AID-JCB15%3E3.0.CO;2-%23
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