Expression vector with DNA of bovine papilloma virus 1 for keratinocyte gene therapy.
Although there are several methods for introducing the genes to keratinocytes in vivo, expression of transgene does not last long enough for effective keratinocyte gene therapy. In this study, we added bovine papilloma virus 1 (BPV) DNA into expression vectors with the lacZ gene driven by metallothionein and keratin 10 promoters, and we transferred them into keratinocytes in vivo using the naked DNA method, and measured beta-gal activity in keratinocytes. The results showed that beta-galactosidase activity of vectors with the BPV DNA was clearly higher than that without the DNA. Moreover, time-course experiment disclosed that the activity of the BPV vector declined at a lower rate than that of the control vector, suggesting this fragment prolonged transgene expression. These results should prove useful for understanding gene regulation in keratinocytes in vivo and for developing potential expression vectors for keratinocyte gene therapy.
Sawamura
D
D
Department of Dermatology, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki, Japan. smartdai@cc.hirosaki-u.ac.jp
Meng
X
X
Ina
S
S
Kon
A
A
Tamai
K
K
Ohe
Y
Y
Hashimoto
I
I
D000818:Animals; D052688:Bovine papillomavirus 1; D002417:Cattle; D004279:DNA, Viral / Q000235:genetics*; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D053550:Keratin-10; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism; D007633:Keratins / Q000235:genetics*; D008668:Metallothionein / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D011919:Rats, Inbred Strains; D011994:Recombinant Proteins / Q000096:biosynthesis; D001616:beta-Galactosidase / Q000235:genetics
D004279:DNA, Viral; D011994:Recombinant Proteins; D053550:Keratin-10; D007633:Keratins; D008668:Metallothionein; D001616:beta-Galactosidase