Enhanced cutaneous gene delivery following intradermal injection of naked DNA in a high ionic strength solution

被引:33
作者
Chesnoy, S [1 ]
Huang, L [1 ]
机构
[1] Univ Pittsburgh, Sch Pharm, Ctr Pharmacogenet, Pittsburgh, PA 15213 USA
关键词
gene therapy; cutaneous gene delivery; naked DNA; intradermal administration;
D O I
10.1006/mthe.2001.0511
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Intradermal injection of naked DNA results in gene transfer to skin cells, but the efficiency of this gene transfer method is relatively low and variable. We have systematically optimized several parameters to obtain reproducible, high-level gene transfer to the mouse skin. Older mice (similar to7 weeks) showed a significant decrease in gene expression compared with younger mice (4-5 weeks old). The composition of the solvent vehicle (electrolyte versus nonelectrolyte) strongly affected gene expression in the skin. A higher level of gene expression was achieved when naked DNA was dissolved in isotonic phosphate buffered saline solution compared with isotonic dextrose solution. Finally, transfection efficiency in older mice was greatly improved by increasing the ionic strength of the solvent vehicle. The improved transfection efficiency was due to an enhanced DNA uptake by the skin cells. Gene transfer was most evident in the subdermal smooth muscle cells and epidermal cells. With the optimized conditions, gene transfer mediated by intradermal injection of naked DNA was comparable in efficiency to electroporation. However, cellular distributions of the gene transfer of the two methods were different.
引用
收藏
页码:57 / 62
页数:6
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