Enhancement of adenovirus-mediated gene transfer into dermal fibroblasts in vitro and in vivo by polyethylene glycol 6000.
Gene therapy directed to the skin requires efficient transfer of the desired gene into cutaneous cells. In this study, we examined several chemical substances present in various ointments by enhancement of virus infectivity. The recombinant adenovirus vector, AxCALacZ, was used to infect dermal fibroblasts with some chemicals both in vitro and in vivo, and the expression of the LacZ gene was determined by X-Gal reaction. As the result, it was shown that PEG 6000 had the highest ability to enhance the exogenous gene expression in cultured fibroblasts with little toxicity. In vivo, it was also demonstrated that fibroblasts in mouse skin were efficiently gene transfer by adenovirus vector and 20% PEG 6000-treatment. These results suggest that this chemical treatment appears to be a simple, safe, convenient, and effective method for facilitating virus-mediated gene therapy in the skin.
Ohsawa
T
T
Department of Molecular Biology, Faculty of Medicine, Tottori University, Yonago, Japan.
Nakamura
T
T
Mihara
M
M
Sato
K
K
D000260:Adenoviruses, Human / Q000235:genetics*; D000328:Adult; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D017931:DNA Primers; D005260:Female; D005347:Fibroblasts / Q000821:virology*; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D011092:Polyethylene Glycols; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000166:cytology / Q000821:virology; D013501:Surface-Active Agents
D017931:DNA Primers; D013501:Surface-Active Agents; D011092:Polyethylene Glycols