In vivo gene transfer method in keratinocyte gene therapy: intradermal injection of DNA complexed with high mobility group-1 protein in rats.
In order to develop a more efficient method of introducing genes into keratinocytes in vivo, we intradermally injected DNA bound to high mobility group 1 protein, thereby taking advantages of the naked DNA and hemagglutinating virus of the Japan-liposome method reported previously. First we performed a gel mobility shift assay, which confirmed DNA binding to high mobility group 1. Then we injected beta-galactosidase expression vector complexed with high mobility group 1 into the rat skin and the activity of sample with the protein was 2-3 times higher than that without the protein as control. Semiquantification of transferred-DNA content using polymerase chain reaction and a time course of transgene expression in keratinocytes suggested that high mobility group 1 protein increased transfer of the DNA from the cytoplasm to the nucleus. Direct injection of the DNA-high-mobility-group-1 complex is a highly efficient method for introducing genes into keratinocytes in connection with gene therapy.
Ina
S
S
Department of Dermatology, Hirosaki University School of Medicine, Japan.
Sawamura
D
D
Meng
X
X
Tamai
K
K
Hanada
K
K
Hashimoto
I
I
D000818:Animals; D004247:DNA / Q000008:administration & dosage*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D006609:High Mobility Group Proteins / Q000008:administration & dosage*; D007271:Injections, Intradermal; D015603:Keratinocytes; D051381:Rats; D011919:Rats, Inbred Strains
D006609:High Mobility Group Proteins; D004247:DNA