Polyplex-mediated gene transfer into human retinal pigment epithelial cells in vitro
Document Type
Journal ArticlePublication Date
1999-12-01Keywords
DNAGene Expression
*Gene Transfer Techniques
Humans
In Vitro Techniques
Pigment Epithelium of Eye
Transgenes
Biochemistry
Cell Biology
Eye Diseases
Ophthalmology
Metadata
Show full item recordAbstract
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-%23DOI
10.1002/(SICI)1097-4644(20000101)76:1%3C153::AID-JCB15%3E3.0.CO;2-%23Permanent Link to this Item
http://hdl.handle.net/20.500.14038/49228PubMed ID
10581009Notes
Medical student Mark P. Hatton participated in this study as part of the Senior Scholars research program at the University of Massachusetts Medical School.
Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1002/(SICI)1097-4644(20000101)76:1%3C153::AID-JCB15%3E3.0.CO;2-%23